[go: up one dir, main page]

WO2020207426A1 - Poche à micropores implantable in vivo, son procédé d'utilisation et son application - Google Patents

Poche à micropores implantable in vivo, son procédé d'utilisation et son application Download PDF

Info

Publication number
WO2020207426A1
WO2020207426A1 PCT/CN2020/083903 CN2020083903W WO2020207426A1 WO 2020207426 A1 WO2020207426 A1 WO 2020207426A1 CN 2020083903 W CN2020083903 W CN 2020083903W WO 2020207426 A1 WO2020207426 A1 WO 2020207426A1
Authority
WO
WIPO (PCT)
Prior art keywords
deficiency
cells
microporous
implantable
bag
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2020/083903
Other languages
English (en)
Chinese (zh)
Inventor
李本尚
陈其民
傅立军
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Childrens Medical Center Affiliated to Shanghai Jiaotong University School of Medicine
Original Assignee
Shanghai Childrens Medical Center Affiliated to Shanghai Jiaotong University School of Medicine
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shanghai Childrens Medical Center Affiliated to Shanghai Jiaotong University School of Medicine filed Critical Shanghai Childrens Medical Center Affiliated to Shanghai Jiaotong University School of Medicine
Publication of WO2020207426A1 publication Critical patent/WO2020207426A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/34—Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/88—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microencapsulation, e.g. using amphiphile liposome vesicle
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention belongs to the field of cell therapy, and specifically relates to an implantable microporous capsule in the body and its use method and application, and specifically relates to an implantable microporous capsule in the body for the treatment of various diseases such as genetic metabolism and Instructions.
  • Genetic metabolic diseases refer to genetic defects in the biosynthesis of certain enzymes, receptors, carriers and membrane pumps composed of polypeptides and (or) proteins that are necessary to maintain normal metabolism of the body, that is, mutations in genes encoding such polypeptides (proteins) The resulting disease, or congenital metabolic defect disease.
  • Inherited metabolic disease is a type of genetic disease with defects in metabolic function, most of which are single-gene genetic diseases, including macromolecular metabolic diseases, including lysosomal storage diseases, mitochondrial diseases, etc.; small molecular metabolic diseases, including amino acids, organic Metabolic genetic diseases such as acid and fatty acid.
  • part of the causes of genetic metabolic diseases are caused by genetic inheritance, and some are caused by acquired genetic mutations. The onset of disease is not only newborns, but covers all ages.
  • the more common genetic metabolic diseases include glycogen storage disease, mucopolysaccharidosis, phenylketonuria, and hepatolenticular degeneration.
  • Inherited metabolic diseases refer to genetic mutations that are inherited from parents, or acquired new mutations, and all or part of an individual's cells have the same genetic mutation.
  • acquired metabolic diseases caused by organ function damage such as diabetes, hyperlipidemia, obesity, etc., are more or less related to the body's genetic susceptibility.
  • Inherited metabolic diseases are the abnormal metabolism of large or small molecules, and some bleeding diseases (such as various hemophilias, thrombosis caused by thrombin deficiency, hypothrombinemia, congenital prothrombin deficiency, etc.) And anemia (such as pyruvate kinase deficiency, 5 ⁇ uridine monophosphate hydrolase deficiency, glucose 6 phosphatase deficiency, adenylate kinase deficiency, etc.) are also caused by mutations in genes encoded by certain enzymes.
  • a few of the above diseases can be alleviated by enzyme replacement therapy, and some diseases can be cured by hematopoietic stem cell transplantation, but more diseases lack effective treatment options, and new treatment concepts and treatment methods are urgently needed.
  • enzyme replacement therapy even if enzyme replacement therapy is adopted, enzyme replacements for many diseases are expensive and require life-long replacement therapy, causing great economic pressure on patients and severely disrupting their normal life.
  • graft-versus-host diseases (GVHD) after transplantation may be very serious and have fatal risks.
  • the expression products of the latter are used to correct body diseases; lentivirus, retrovirus, adenovirus, adeno-associated virus, transposon, plasmid
  • Various methods such as direct transfection, protein transfection, electrotransformation, etc. modify the genome, transcription, post-transcriptional modification, translation, etc., so that autologous cells can express specific genes; through the use of genetically normal induced pluripotent stem cells Induced expression of specific tissue cell types by (IPS) technology, and achieves the purpose of alternative treatment after transfusion into the human body.
  • IPS genetically normal induced pluripotent stem cells Induced expression of specific tissue cell types by
  • Proteins, etc. allowing human immune cells to recognize the modified gene-edited cells, causing them to not be able to function for a long time in the body.
  • these modified cells may cause the insertion and inactivation of tumor suppressor genes in the genome or the overexpression of oncogenes, etc., so there is a long-term carcinogenic risk.
  • foreign elements are inserted into the genome of these cells, which may cause problems with the stability of the genome, and the number of divisions and passages of the corresponding cells may be reduced, resulting in cell senescence and loss of functional activity. Based on this, there is an urgent clinical need for a technology that can last for a long time without being recognized and attacked by autoimmune cells to solve the above problems.
  • the technical problem to be solved by the present invention is to overcome the shortcomings of the prior art and provide an implantable microporous capsule that can last for a long time and is not recognized and attacked by autoimmune cells.
  • the implantable microporous capsule The bag is composed of biological tissues or materials with good compatibility, and its pore size is not greater than 0.45 microns, preferably not greater than 0.22 microns. It can exchange proteins and other biological macromolecules with the body, but it can prevent immune cells from entering the sac, and has immunity The role of a shelter. Through various enzymes and other protein molecules that the body lacks secreted by cells in the capsular bag, it has a therapeutic effect on various diseases such as genetic metabolism.
  • the first object of the present invention is to provide a microporous capsule that can be implanted in the body.
  • the microporous capsule includes a bag-like structure that is made of at least one layer of biological tissue or a membrane made of a tissue compatible material.
  • the membrane is surrounded by micropores, and the diameter of the micropores is not greater than 0.45 microns.
  • the diameter of the micropores is not greater than 0.22 microns.
  • the present invention proposes a new concept of implantable immune shelters for humans, by providing a kind of implantable microporous capsules that can last for a long time and are not recognized and attacked by autoimmune cells.
  • the bag can be taken from normal human tissues, or made of biological tissues or materials with good compatibility. It can be retained in the human body for many years without causing the decline of body cells and tissue functions, and the tissues will not undergo inflammatory reactions, carcinogenesis, and Rejection.
  • the capsular bag can be the source of normal human tissues, or it can be composed of one or more layers of biological tissues or materials with good compatibility.
  • the diameter of the micropores is not more than 0.22 microns, which can effectively prevent the cells and bacteria in the capsular bag from passing through
  • the micropores on the capsular bag enter the blood circulation, which can also effectively prevent immune cells in the blood from entering the capsular bag to reject the cells in the capsular bag, thereby playing a physical shielding role and forming an implantable immune shelter in the body.
  • Immune shelter refers to the part that immune cells cannot reach. Therefore, the body's immune cells cannot produce cytotoxic effects on the gene-edited autologous cells or non-autologous foreign cells in the sac, and the gene-edited cells in the sac Autologous cells or non-autologous foreign cells can't have an effect on the body.
  • antibodies and complement components in body fluids can freely pass through the micropores, these antibodies and complement components need to rely on lymphocytes, monocyte-macrophages, etc. to function, and because the latter cannot enter the microporous pockets, It will not cause an immune attack on the cells in the sac.
  • the cellular components in the microporous sac may produce foreign antigens for the body due to cell necrosis, these antigens will only stimulate the body to produce corresponding antibodies or specific reactive lymphocytes, the latter as long as the body does not The cross-reactivity will not cause harm to the human body.
  • the bag-like structure includes a multilayer film, each film has micropores, and the preparation materials of each film are the same or different;
  • the material for preparing the membrane includes one or more of biological tissues, titanium alloys, natural polymer materials, synthetic polymer materials, nano materials, and human-derived materials.
  • the bag-like structure includes a multilayer film, and the toughness of the film on the outer layer is not less than the toughness of the film on the inner layer; preferably, the film on the inner layer is made of nano materials and is located on the outer layer.
  • the membrane is made of titanium alloy material.
  • the microporous pouch of the present application is made of normal human tissues, or biological tissues or materials with good compatibility. It can be retained in the human body for many years without causing the function of the body's cells and tissues to decline. Inflammatory reaction, cancer and rejection. Biological tissues or materials with good compatibility can choose titanium alloys, natural polymer materials, synthetic polymer materials, nano materials, etc. These materials have strong toughness and strength, and can withstand the impact of perennial blood flow. Fracture and breakage.
  • These materials can be composed of one layer or multiple layers, of which the outermost layer material needs to provide a certain strength, preferably titanium alloy material as the outermost layer, to protect the inner layer material; the inner layer can choose the above-mentioned materials, preferably Nanomaterials provide a broad surface area for cell growth in the capsular bag.
  • the bag-like structure has a cavity inside, and the cavity is filled with cells that can secrete drugs, and the microporous sac forms an immune shelter that cannot be reached by immune cells.
  • Cells include human cells and non-human cells. Human cells can be taken from autologous or exogenous healthy human bodies. Different donor cell sources can be selected according to specific disease needs.
  • the cells include autologous cells and/or exogenous cells, which can secrete proteins and/or small molecules for treating diseases; the implantable microporous pouches in the present application contain unmodified or gene-edited
  • the modified autologous cells or non-autologous exogenous cells can produce various drugs such as proteins and small molecules that have therapeutic value for patients' diseases.
  • the cells include unmodified cells, and/or gene-edited cells.
  • the gene editing technology used in this application includes the use of TALEN, ZFN, CRISPR and other technologies to edit the genome of autologous cells to efficiently express mutation-free genes related to genetic defects.
  • the expression products of the latter can be used to correct body diseases; including Various methods such as lentivirus, retrovirus, adenovirus, adeno-associated virus, transposon, plasmid direct transfection, protein transfection, electrotransformation, etc. transform the genome, transcription, post-transcriptional modification, translation, etc., for all purposes.
  • IPS induced pluripotent stem cell
  • the autologous cells include cells derived from various tissues of the body; the exogenous cells include cells derived from various tissues of the body;
  • autologous cells include bone marrow hematopoietic stem cells, mesenchymal stromal cells, fibroblasts, epithelial cells, muscle cells, adipocytes, adipose stem cells, endothelial cells, cardiomyocytes, neurons and glial cells, skin cell;
  • the exogenous cells include non-autologous human cells, non-human cells, cell lines, and engineered cells;
  • non-autologous human-derived cells include bone marrow hematopoietic stem cells, mesenchymal stromal cells, fibroblasts, epithelial cells, muscle cells, adipocytes, adipose stem cells, endothelial cells, cardiomyocytes, neurons and glial cells, skin Cells; non-human cells include mammalian cells, insect cells, plasmodium, and bacteria.
  • the microporous capsule further includes a pipe structure communicating with the cavity, through which substances are added to or extracted from the microporous capsule.
  • the microporous pouch is a complete whole, and the diameter of the micropores on the biological tissue or the material with good compatibility is not more than 0.22 micrometer (M).
  • M micrometer
  • biological macromolecules such as proteins can pass freely, but cells and bacteria cannot enter and exit through the micropores, which can effectively prevent cells and bacteria in the pouch from entering the blood circulation through the micropores on the pouch. It effectively prevents immune cells in the blood from entering the pouch and repelling the cells in the pouch, thereby playing a physical shielding role and forming an implantable immune shelter in the body.
  • the intact capsular bag is connected to a tube, which is also composed of materials with good tissue compatibility.
  • One end of the tube is seamlessly connected to the microporous capsular bag, and the other end can be fixed under the skin or the skin surface.
  • the implantable microporous capsule in the body can be fixed to the subcutaneous, skin surface, etc. through a connected tube, which facilitates the observation and addition or extraction of cell components in the microporous capsule.
  • the pipeline structure is hollow inside and made of biological tissue or tissue compatible material
  • the pipe structure and the bag-like structure are made of the same or different materials;
  • one end of the pipe structure is seamlessly connected with the bag-like structure, and the other end is fixed under the skin or the skin surface.
  • the second object of the present invention is to provide a method for using the implantable microporous capsule in the body as described in any one or several combinations of the above, wherein the microporous capsule is completely implanted in the body or fixed To the subcutaneous or skin surface;
  • the bag-like structure of the microporous capsule is completely implanted in the body, and the free end of the tube structure is fixed to the subcutaneous or skin surface.
  • the cell components in the implantable microporous capsule of the present application can be sampled and tested through a pipe connected to the capsule, which is convenient for understanding the growth of cells in the capsule.
  • disease dynamics can be monitored by detecting the concentration of corresponding disease proteins and small molecule drugs in the blood.
  • the microporous capsule is implanted into at least one of the heart, abdominal cavity, thoracic cavity, bone marrow cavity, spleen, liver, subarachnoid space, cerebral ventricle, and joint cavity;
  • the microporous capsular bag is implanted into the heart through the upper arm's main vein-brachial vein-axillary vein-subclavian vein-brachiocephalic vein-superior vena cava-right atrium.
  • the specific implantation site is not limited to the above-mentioned path and final location, and implantation can be performed according to specific diseases and needs.
  • the third object of the present invention is to provide an application of the implantable microporous pouch in the body as described in any one or several combinations of the above in the treatment of genetic metabolic diseases.
  • the genetic metabolic diseases include congenital metabolic diseases. Acquired metabolic diseases and hemorrhagic diseases caused by low organ function;
  • the congenital metabolic diseases include glycogen storage disease, mucopolysaccharidosis, congenital abnormal glycosylation, hyperlipoproteinemia, leukodystrophy, neuronal wax-like lipofuscin deposition Disease, hepatolenticular degeneration, aromatic amino acid decarboxylase deficiency, cellular peroxidase deficiency, fructose-1,6-diphosphate kinase deficiency, hemolytic anemia caused by G6PD deficiency, Gaucher disease, Fab Li disease, ganglioside storage disease, mucolipid storage disease, hypoxanthine guanine phosphoribosyl transferase deficiency, hyaluronidase deficiency, lactose intolerance, myeloperoxidase deficiency, no Beta lipoproteinemia, pyruvate carboxylase deficiency, lactic acidemia, phosphoglycerate dehydrogenase defic
  • acquired metabolic diseases caused by low organ function include various types of diabetes, obesity, diabetes insipidus, hyperlipidemia, hypoparathyroidism, hypothyroidism, hypofunction of adrenal cortex, and shortness;
  • the bleeding disorder includes hemophilia A, hemophilia B, hemophilia C, von Willebrand disease, thrombosis caused by thrombin deficiency, hypothrombinemia, and congenital prothrombin deficiency , Prothrombin dysplasia, thrombosis due to thrombin deficiency, hereditary factor V deficiency, thrombosis, thrombocytopenia caused by X-linked factor IX deficiency, factor XII deficiency, hereditary factor XIIIA deficiency Syndrome, hereditary factor XIIIB deficiency, high molecular weight kininogen deficiency, combined factor V and factor VIII deficiency, congenital anemia/hypofibrinogenemia, plasminogen activator inhibitor 1 Deficiency, ⁇ 2-plasmin inhibitor deficiency, plasminogen deficiency, pyruvate kinase deficiency, 5-ur
  • the present invention has the following beneficial effects compared with the prior art:
  • the present invention provides an implantable microporous pouch that can last for a long time and is not recognized and attacked by autoimmune cells.
  • the implantable microporous pouch is composed of biological tissues or materials with good compatibility.
  • the pore size of the micropore is not greater than 0.45 microns, preferably 0.22 microns, which can exchange proteins and other biological macromolecules with the body, and at the same time can prevent immune cells from entering the microporous sac, which has the function of immune shelter of the body.
  • Various enzymes and other protein molecules lacking in the body secreted by the cells in the microporous pouch have a therapeutic effect on various diseases such as genetic metabolism.
  • Fig. 1 is a schematic diagram of the position selection of the microporous capsule implanted in the human body according to the present invention; specifically, the implantable microporous capsule in the body can pass through the important vein of the right upper arm—brachial vein—axillary vein—subclavian vein— Brachiocephalic vein-superior vena cava-right atrium; it can also be implanted in any part of the body including abdominal cavity, thoracic cavity, bone marrow cavity, spleen, liver, subarachnoid space, cerebral ventricle, joint cavity, etc.;
  • Figure 2 is a schematic diagram of the structure of the microporous capsule of the present invention; in the figure: 1 bag-like structure, 2 pipe structure;
  • Figure 3 is the outer structure of the microporous capsule implanted in the body; it is composed of a titanium alloy mesh with a mesh size of 10-100 ⁇ M, which mainly provides support for the inner biofilm structure.
  • Figure 4 is the internal structure of the implantable microporous capsule in vivo; the internal layer is composed of an organic biofilm synthesized in vitro, which is induced to differentiate into fibroblasts from human peripheral blood mononuclear cells, and then express a variety of adhesion molecules It is formed by cross-linking each other in multiple layers, with relatively strong toughness;
  • Figure 5 is a method for making an implantable microporous capsule in vivo; the biofilm is folded into a bag shape, and the mouth of the bag is gently tied with surgical thread, and a certain amount of induced cardiomyocytes transfected with foreign genes is to be injected After the capsular bag, the surgical thread is ligated to close the bag mouth.
  • Figure 6 shows the steps of inducing cardiomyocytes in vitro; specifically, the steps of inducing peripheral blood mononuclear cells into cardiomyocytes in vitro;
  • Figure 7 shows the cardiomyocytes successfully induced in vitro; after the cardiomyocytes are induced in vitro, rhythmic beating appears in the cardiomyocytes, indicating the successful induction;
  • Figure 8 is the identification of induced cardiomyocytes in vitro; the detection of a variety of cardiomyocyte specific antigens confirmed that cardiomyocytes were successfully induced;
  • Figure 9 is the construction of the human insulin expression vector pWPXL-Insulin-2A-myc; the vector pWPXL is selected as the target vector, and the human insulin full-length CDS sequence and the myc antigen structure linked by the 2A sequence are expressed and linked into the above-mentioned vector, and the corresponding The virus backbone was co-transfected into human 293T cells to obtain the corresponding lentivirus.
  • Figure 10 is the detection of Insulin expression in Hela and induced cardiomyocytes; lentivirus overexpressing human insulin infects human cervical cancer cell line-Hela and induced cardiomyocytes, and then expresses insulin through western blot after total protein extraction Level detection, the first track on the left in Figure 10 is the expression level of insulin in Hela cells, the middle is the expression level of insulin in the induced cardiomyocytes, and the right is the level of insulin expression in the control transfected virus-laden cells. It can be seen from the figure that the expression of specific insulin can be seen in both Hela and the induced cardiomyocytes.
  • this embodiment provides a microporous capsule implantable in the body.
  • the microporous capsule includes a bag-like structure composed of at least one layer of biological tissue or tissue compatibility.
  • the material is surrounded by a film with micropores, and the diameter of the micropores is not greater than 0.45 microns. In a preferred solution, the diameter of the micropores is not greater than 0.22 microns.
  • This program proposes a new concept of implantable immune shelters for humans, by providing a kind of implantable microporous capsules that can last for a long time and are not recognized and attacked by autoimmune cells.
  • the bag can be taken from normal human tissues, or made of biological tissues or materials with good compatibility. It can be retained in the human body for many years without causing the decline of body cells and tissue functions, and the tissues will not undergo inflammatory reactions, carcinogenesis, and Rejection.
  • the material of the microporous pouch can be from normal human tissues, or it can be composed of one or more layers of biological tissue or materials with good compatibility.
  • the diameter of the micropores is not greater than 0.45 microns, preferably less than or equal to 0.22 microns, which can be effective Prevent cells and bacteria in the capsular bag from entering the blood circulation through the micropores on the capsular bag. It can also effectively prevent immune cells in the blood from entering the capsular bag and repelling the cells in the capsular bag, thereby playing a physical shielding role and forming an in vivo implant. Access to sexual immunity shelters.
  • Immune shelter refers to the part that immune cells cannot reach. Therefore, the body's immune cells cannot produce cytotoxic effects on the gene-edited autologous cells or non-autologous foreign cells in the sac, and the gene-edited cells in the sac Autologous cells or non-autologous foreign cells can't have an effect on the body.
  • antibodies and complement components in body fluids can freely pass through the micropores, these antibodies and complement components need to rely on lymphocytes, monocyte-macrophages, etc. to function, and because the latter cannot enter the microporous pockets, It will not cause an immune attack on the cells in the sac.
  • the cellular components in the microporous sac may produce foreign antigens for the body due to cell necrosis, these antigens will only stimulate the body to produce corresponding antibodies or specific reactive lymphocytes, the latter as long as the body does not The cross-reactivity will not cause harm to the human body.
  • the bag-like structure includes a multilayer film, each film has micropores, and the preparation materials of each film are the same or different;
  • the preparation materials of the membrane include one or more of biological tissues, titanium alloys, natural polymer materials, synthetic polymer materials, nano materials, and materials derived from human bodies.
  • the bag-like structure includes a multilayer film, and the toughness of the film on the outer layer is not less than the toughness of the film on the inner layer; preferably, the film on the inner layer is made of nanomaterials, and the film on the outer layer is made of titanium Made of alloy material.
  • the microporous pouch of the present application is made of normal human tissues, or biological tissues or materials with good compatibility. It can be retained in the human body for many years without causing the function of the body's cells and tissues to decline. Inflammatory reaction, cancer and rejection. Biological tissues or materials with good compatibility can choose titanium alloys, natural polymer materials, synthetic polymer materials, nano materials, etc. These materials have strong toughness and strength, and can withstand the impact of perennial blood flow. Fracture and breakage.
  • These materials can be composed of one layer or multiple layers, of which the outermost layer material needs to provide a certain strength, preferably titanium alloy material as the outermost layer, to protect the inner layer material; the inner layer can choose the above-mentioned materials, preferably Nanomaterials provide a broad surface area for cell growth in the capsular bag.
  • the bag-like structure has a cavity inside, and the cavity is filled with cells that can secrete drugs, and the microporous sac forms an immune shelter that cannot be reached by immune cells.
  • the bag-like structure is surrounded by at least one layer of biological tissue or a membrane made of a tissue compatible material, and has an opening that can be used to fill the cavity with cells that can secrete drugs.
  • the microporous capsule does not have a pipe structure, but only has a bag-like structure, the cavity can be filled with drug-secreting cells, the opening can be closed, and then implanted into the human body.
  • the cells include human cells and non-human cells.
  • Human cells can be taken from autologous or exogenous healthy humans, and different donor cell sources can be selected according to specific disease requirements.
  • the cells include autologous cells and/or exogenous cells, which can secrete proteins and/or small molecules for treating diseases; the implantable microporous pouches in the present application contain unmodified or modified by different methods such as gene editing Autologous cells or non-autologous exogenous cells, these cells can produce a variety of drugs such as proteins and small molecules that have therapeutic value for patients' diseases.
  • the cells include unmodified cells, and/or gene-edited cells.
  • the gene editing technology used in this application includes the use of TALEN, ZFN, CRISPR and other technologies to edit the genome of autologous cells to efficiently express mutation-free genes related to genetic defects.
  • the expression products of the latter can be used to correct body diseases; including Various methods such as lentivirus, retrovirus, adenovirus, adeno-associated virus, transposon, plasmid direct transfection, protein transfection, electrotransformation, etc. transform the genome, transcription, post-transcriptional modification, translation, etc., for all purposes.
  • IPS induced pluripotent stem cell
  • the autologous cells include cells derived from various tissues of the body; the exogenous cells include cells derived from various tissues of the body;
  • Autologous cells include bone marrow hematopoietic stem cells, mesenchymal stromal cells, fibroblasts, epithelial cells, muscle cells, adipocytes, adipose stem cells, endothelial cells, cardiomyocytes, neurons and glial cells, and skin cells taken from the body;
  • the exogenous cells include non-autologous human cells, non-human cells, cell lines, and engineered cells;
  • Non-autologous human-derived cells include bone marrow hematopoietic stem cells, mesenchymal stromal cells, fibroblasts, epithelial cells, muscle cells, adipocytes, adipose stem cells, endothelial cells, cardiomyocytes, neurons and glial cells, and skin cells; Human-derived cells include mammalian cells, insect cells, plasmodium, and bacteria.
  • the microporous capsule includes a bag-like structure and a pipe structure.
  • the bag-like structure has a cavity inside, and the pipe structure communicates with the cavity through the pipe structure. Add substances to or extract substances from the bag-like structure.
  • the microporous capsule is a complete whole, and the diameter of the micropores on the biological tissue or the material with good compatibility is not more than 0.22 micrometers. Below this scale, biological macromolecules such as proteins can pass freely, but cells and bacteria cannot enter and exit through the micropores, which can effectively prevent cells and bacteria in the pouch from entering the blood circulation through the micropores on the pouch. It effectively prevents immune cells in the blood from entering the pouch and repelling the cells in the pouch, thereby playing a physical shielding role and forming an implantable immune shelter in the body.
  • the intact capsular bag is connected to a tube, which is also composed of materials with good tissue compatibility.
  • One end of the tube is seamlessly connected to the microporous capsular bag, and the other end can be fixed under the skin or the skin surface.
  • the implantable microporous capsule in the body can be fixed to the subcutaneous, skin surface, etc. through a connected tube, which facilitates the observation and addition or extraction of cell components in the microporous capsule.
  • the pipeline structure is hollow inside and made of biological tissue or tissue compatible material
  • the pipe structure and the bag-like structure are made of the same or different materials
  • One end of the pipe structure is seamlessly connected with the bag-like structure, and the other end is fixed under the skin or the skin surface.
  • this solution also provides a method for using the implantable microporous capsule in the body as described in any one or several combinations of the above.
  • the microporous capsule is completely implanted in the body or fixed under the skin. Or the skin surface; the bag-like structure of the microporous capsule is completely implanted in the body, and the free end of the tube structure is fixed to the subcutaneous or skin surface.
  • the cell components in the implantable microporous capsule of the present application can be sampled and tested through a pipe connected to the capsule, which is convenient for understanding the growth of cells in the capsule.
  • disease dynamics can be monitored by detecting the concentration of corresponding disease proteins and small molecule drugs in the blood, and ingredients can also be added to the microporous capsule.
  • the microporous capsule is implanted in at least one of the heart, abdominal cavity, thoracic cavity, bone marrow cavity, spleen, liver, subarachnoid space, cerebral ventricle, and joint cavity;
  • the microporous capsule is implanted into the heart through the right upper arm's main vein-brachial vein-axillary vein-subclavian vein-brachiocephalic vein-superior vena cava-right atrium.
  • the specific implantation site is not limited to the above-mentioned path and final location, and implantation can be performed according to specific diseases and needs.
  • this solution also provides an application of implantable microporous pouches in the body as described in any one or several combination solutions in the treatment of genetic metabolic diseases.
  • Genetic metabolic diseases include congenital metabolic diseases, organs Acquired metabolic diseases and bleeding diseases caused by hypofunction;
  • the congenital metabolic diseases include glycogen storage disease, mucopolysaccharidosis, congenital abnormal glycosylation, hyperlipoproteinemia, leukodystrophy, neuronal wax-like lipofuscin deposition Disease, hepatolenticular degeneration, aromatic amino acid decarboxylase deficiency, cellular peroxidase deficiency, fructose-1,6-diphosphate kinase deficiency, hemolytic anemia caused by G6PD deficiency, Gaucher disease, Fab Li disease, ganglioside storage disease, mucolipid storage disease, hypoxanthine guanine phosphoribosyl transferase deficiency, hyaluronidase deficiency, lactose intolerance, myeloperoxidase deficiency, no Beta lipoproteinemia, pyruvate carboxylase deficiency, lactic acidemia, phosphoglycerate dehydrogenase defic
  • acquired metabolic diseases caused by low organ function include various types of diabetes, obesity, diabetes insipidus, hyperlipidemia, hypoparathyroidism, hypothyroidism, hypofunction of adrenal cortex, and shortness;
  • the bleeding disorder includes hemophilia A, hemophilia B, hemophilia C, von Willebrand disease, thrombosis caused by thrombin deficiency, hypothrombinemia, and congenital prothrombin deficiency , Prothrombin dysplasia, thrombosis due to thrombin deficiency, hereditary factor V deficiency, thrombosis, thrombocytopenia caused by X-linked factor IX deficiency, factor XII deficiency, hereditary factor XIIIA deficiency Syndrome, hereditary factor XIIIB deficiency, high molecular weight kininogen deficiency, combined factor V and factor VIII deficiency, congenital anemia/hypofibrinogenemia, plasminogen activator inhibitor 1 Deficiency, ⁇ 2-plasmin inhibitor deficiency, plasminogen deficiency, pyruvate kinase deficiency, 5-ur
  • Example 1 An implementation of the microporous capsule
  • the microporous pouch is composed of two layers, and the outer layer is composed of a titanium alloy mesh (Figure 3), which provides a physical support for the pouch.
  • the mesh size of the titanium alloy mesh is 10-100 ⁇ M, which allows various components in the blood to pass through the mesh freely.
  • the inner layer is composed of an organic biofilm synthesized in vitro.
  • the biofilm is made from human peripheral blood mononuclear cells induced to differentiate into fibroblasts, and then multi-layered and cross-linked after expressing a variety of adhesion molecules. It is tough, without blood vessels and nerves, and contains a lot of collagen, forming a dense structure.
  • the biofilm has no immunogenicity and can produce immune isolation through the cell structure of the biofilm species, and is a key component of the microporous pouch.
  • the image of the organic biofilm in the culture medium is shown in Figure 4.
  • the organic biofilm is first folded into a bag shape, and the mouth of the pouch is gently tied with surgical thread, and a certain number of induced cardiomyocytes transfected with exogenous genes will be injected into the pouch.
  • the surgical thread is ligated to close the bag mouth (as shown in Figure 5). Then use titanium alloy mesh to wrap the biofilm capsule to provide physical support.
  • the biofilm After the above-mentioned capsular bag is placed in the human body, the biofilm has a translucent effect. Although there is no neurovascular distribution, the body fluid around the membrane can provide its growth, and small molecules such as water, inorganic salts, and large molecules such as plasma proteins can penetrate Permeable biomembrane can nourish the cells in the sac, but the cells in the sac cannot pass through the biomembrane, so the biological sac can play a role in immune isolation.
  • cardiomyocytes have the characteristics of long life span, no proliferation and division, and ability to pulsate autonomously, they are the most suitable genetic engineering cell carrier for expressing foreign genes in microporous pouches.
  • Cardiomyocytes can obtain induced pluripotent stem cells (iPS) by transfecting the four transcription factors Oct3/4, Sox2, c-Myc and Klf4 with monocytes in peripheral blood in vitro, and stimulated by a series of exogenous small molecules Obtained (the process is shown in Figure 6).
  • peripheral blood mononuclear cells were cultured in vitro, transfected with lentiviral vectors inserted into the four transcription factors Oct3/4, Sox2, c-Myc and Klf4, and then cultured to obtain iPS.
  • the differentiation medium I is added, and the culture is continued for 48 hours. Subsequently, the culture medium I was removed, and the culture medium III was added to continue the culture for 24 hours. Then remove the culture solution III and add the culture solution II for another 48 hours. Then remove the medium II and add the medium III to culture for 48 hours. Use medium III to continuously change the medium until the cell beating occurs, at which time the cardiomyocytes are successfully induced (as shown in Figure 7).
  • Troponin T Troponin T
  • Troponin I troponin I
  • Alpha Actin anti-human alpha actinin
  • Diabetes is a group of metabolic diseases characterized by hyperglycemia, which is mainly caused by insulin secretion defects or impaired biological effects, or both.
  • the long-term high blood sugar in diabetes causes chronic damage and dysfunction of various tissues, especially the eyes, kidneys, heart, blood vessels, and nerves.
  • oral hypoglycemic drugs and injection of animal insulin human insulin and insulin analogs are the main measures.
  • the human insulin gene contains 110 amino acids.
  • the biologically active insulin molecule is a small protein protein containing 51 amino acids. It is composed of the above-mentioned A chain peptide (21 amino acids) and B chain peptide (30 amino acids) through disulfide bonds.
  • pancreatic islet B cells first synthesize a large molecule of proinsulin, which is processed into 86 peptide proinsulin after the signal peptide is excised, and the latter is hydrolyzed into insulin (A chain peptide and B chain peptide) and connecting peptide (C Chain peptide).
  • the human insulin gene sequence with restriction sites and the specific membrane protein expression tag (myc) were inserted into the lentiviral vector pWPXL by means of gene synthesis to construct the vector pWPXL-Insulin-2A-myc ( Figure 9). After co-transfecting 293T cells with VSVG and other backbone vectors, the lentivirus was prepared, concentrated and purified.
  • the virus was transfected into the human cervical cancer cell line-Hela and the cardiomyocytes induced in vitro. After 48 hours, the cells were collected for protein extraction. After the protein was quantified, the protein was electrophoresed in a 15% gel. Then transfer the protein band from the polyacrylamide gel to the surface of the PVDF membrane under a voltage of 100V. After one hour of electroporation, it will be combined with the specific antibody. The secondary antibody and the coloring agent are colored on the film. It can be seen that the molecular weight is about 10kd. The left and right strips are shown in Figure 10.
  • the first track on the left is the expression level of insulin in Hela cells
  • the middle is the expression level of insulin in the induced cardiomyocytes
  • the right is the expression level of insulin in the control transfected virus-laden cells. The expression of specific insulin was seen in Hela and induced cardiomyocytes.
  • the modified cardiomyocytes that can express insulin are injected into the capsular bag in Example 1, the surgical thread is ligated to close the bag mouth, and then the biofilm capsular bag is wrapped with titanium alloy mesh and placed in a diabetic patient. Cardiomyocytes continue to produce insulin to make up for the lack of insulin and achieve the purpose of treating diabetes.
  • cells that can secrete corresponding drugs are constructed, injected into the capsular bag in Example 1, and placed in the patient's body to achieve the corresponding therapeutic purposes, which will not be described here.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Diabetes (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Epidemiology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Zoology (AREA)
  • Obesity (AREA)
  • Genetics & Genomics (AREA)
  • Developmental Biology & Embryology (AREA)
  • Vascular Medicine (AREA)
  • Cardiology (AREA)
  • Emergency Medicine (AREA)
  • General Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Endocrinology (AREA)
  • Immunology (AREA)
  • Virology (AREA)
  • Botany (AREA)
  • Inorganic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Neurosurgery (AREA)
  • Plant Pathology (AREA)

Abstract

L'invention concerne une poche à micropores implantable in vivo, un procédé d'utilisation correspondant et une application associée. La poche à micropores comprend une structure de poche (1), la structure de poche (1) étant entourée par un film constitué d'au moins une couche d'un tissu biologique ou d'un matériau compatible avec un tissu, des micropores étant formés dans le film, et le diamètre des micropores n'étant pas supérieur à 0,45 micromètres. L'intérieur de la structure de poche (1) est pourvu d'une cavité, et des cellules aptes à sécréter un médicament pour un trouble métabolique hérité correspondant sont contenues dans la cavité. En utilisant une fonction de protection physique de la taille des pores de la poche à micropores, une attaque, à partir d'un facteur immunitaire de l'organisme, contre des cellules autologues ou des cellules exogènes non autologues qui ne sont pas modifiées ou qui sont génétiquement éditées et modifiées dans la poche à micropores peut être efficacement isolée, et également par l'implantation de la poche à micropores dans une oreillette droite, une cavité médullaire ou une autre partie d'un patient, les cellules dans la poche à micropores peuvent être suffisamment alimentées en sang, de telle sorte que les cellules dans la poche à micropores génèrent en continu des substances de substitution requises par l'organisme.
PCT/CN2020/083903 2019-04-10 2020-04-09 Poche à micropores implantable in vivo, son procédé d'utilisation et son application Ceased WO2020207426A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201910284644 2019-04-10
CN201910284644.0 2019-04-10

Publications (1)

Publication Number Publication Date
WO2020207426A1 true WO2020207426A1 (fr) 2020-10-15

Family

ID=72750967

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2020/083903 Ceased WO2020207426A1 (fr) 2019-04-10 2020-04-09 Poche à micropores implantable in vivo, son procédé d'utilisation et son application

Country Status (2)

Country Link
CN (1) CN111840207A (fr)
WO (1) WO2020207426A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114600835B (zh) * 2022-03-08 2023-01-24 郑羽 一种实验室pdx模型建立辅助系统及其控制方法
CN115487357A (zh) * 2022-08-18 2022-12-20 华南理工大学 一种孔径可控的免疫隔离细胞封装囊袋及其制备方法与应用

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102361971A (zh) * 2009-01-23 2012-02-22 Ns基因公司 改善的细胞系以及在胶囊化细胞生物递送中的用途
CN104922684A (zh) * 2015-05-28 2015-09-23 杭州优玛达生物科技有限公司 天然来源和/或自组装技术得到的生物膜、具有生物膜性的闭合结构或细胞区室及其制备方法和应用
CN106039421A (zh) * 2015-04-07 2016-10-26 四川蓝光英诺生物科技股份有限公司 一种包含内皮细胞的生物砖及其用途
CN107802890A (zh) * 2010-01-14 2018-03-16 奥加诺吉尼西斯公司 生物工程化组织构建物及其制备和使用方法
CN108289942A (zh) * 2015-09-25 2018-07-17 迈斯免疫公司 用生产免疫调节剂的免疫-分离的细胞接种疫苗
CN109071597A (zh) * 2016-03-15 2018-12-21 科迪艾克生物科学公司 治疗性膜囊泡
EP3428264A1 (fr) * 2017-07-12 2019-01-16 Defymed Poche non pliable pour former un organe artificiel implantable

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1206044A (zh) * 1997-07-21 1999-01-27 德国赫彻斯特马里奥罗塞尔有限公司 遗传修饰的细胞和它们在预防或治疗疾病中的用途
CN102399740A (zh) * 2010-09-19 2012-04-04 林雄斌 使人的体细胞逆向分化产生自体胰腺干细胞和自体胰岛的方法、试剂盒及其应用

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102361971A (zh) * 2009-01-23 2012-02-22 Ns基因公司 改善的细胞系以及在胶囊化细胞生物递送中的用途
CN107802890A (zh) * 2010-01-14 2018-03-16 奥加诺吉尼西斯公司 生物工程化组织构建物及其制备和使用方法
CN106039421A (zh) * 2015-04-07 2016-10-26 四川蓝光英诺生物科技股份有限公司 一种包含内皮细胞的生物砖及其用途
CN104922684A (zh) * 2015-05-28 2015-09-23 杭州优玛达生物科技有限公司 天然来源和/或自组装技术得到的生物膜、具有生物膜性的闭合结构或细胞区室及其制备方法和应用
CN108289942A (zh) * 2015-09-25 2018-07-17 迈斯免疫公司 用生产免疫调节剂的免疫-分离的细胞接种疫苗
CN109071597A (zh) * 2016-03-15 2018-12-21 科迪艾克生物科学公司 治疗性膜囊泡
EP3428264A1 (fr) * 2017-07-12 2019-01-16 Defymed Poche non pliable pour former un organe artificiel implantable

Also Published As

Publication number Publication date
CN111840207A (zh) 2020-10-30

Similar Documents

Publication Publication Date Title
ES2706401T3 (es) Estructura saquiforme con actividad paracrina y métodos para su preparación
EP1007631B2 (fr) Regeneration du muscle cardiaque a l'aide de cellules souche mesenchymateuses
US8071085B2 (en) Primary cultured adipocytes for gene therapy
US9913865B2 (en) Induced pacemaker and Purkinje cells from adult stem cells
JP7538210B2 (ja) がん細胞療法用のｐ２１発現単球
CN107427535A (zh) 经修饰的血凝块
ES2291588T3 (es) Celulas estromales aisladas y metodos de uso e las mismas.
WO2020207426A1 (fr) Poche à micropores implantable in vivo, son procédé d'utilisation et son application
CN110291190A (zh) 骨骼肌细胞及其诱导方法
KR101815080B1 (ko) 췌장소도세포 및 엘라스틴 유사 인공 세포외 기질을 포함하는 당뇨병 치료용 약학적 조성물
Matsui Endothelial progenitor cell-based therapy for hemophilia A
JP3609395B2 (ja) 哺乳類の不死化肝臓細胞
CN114854693B (zh) 组织工程化传导束传导速度的调控方法及应用
EP4461302A1 (fr) Composition pour le traitement d'un trouble de la fonction hépatique
US20250090587A1 (en) Platelet-derived mitochondria-containing extracellular vesicles for use in the treatment of ocular disorders
CN114058591B (zh) 一种重组间充质干细胞及其应用
CN118403213A (zh) 一种SIRT1基因修饰BMSCs的水凝胶涂层多孔镁合金新型复合材料
Coco-Martin et al. Cell therapy replacement for retinal and optic nerve diseases: cell sources, clinical trials and challenges
US20220016219A1 (en) Means and Methods to Treat Diabetes
CA2330172A1 (fr) Implant prothetique et techniques d'utilisation aux fins de l'expression de genes therapeutiques
CN116531405A (zh) 脐带间充质干细胞源小细胞外囊泡在制备治疗二型糖尿病药物中的应用
WO2022120214A1 (fr) Traitement du diabète
CN116421729A (zh) 脂肪细胞增强结合蛋白1在治疗组织纤维化中的功能及应用
CN116622638A (zh) 一种lamp2b/vwf-a3区域共修饰的外泌体及其制备方法和应用
WO2021248408A1 (fr) Application d'arn circulaire à la régulation et au contrôle de la prolifération des cellules b des îlots pancréatiques

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20787434

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20787434

Country of ref document: EP

Kind code of ref document: A1