[go: up one dir, main page]

WO2019162790A1 - Procédé de fabrication d'un produit de bio-ingénierie utilisant la bio-impression tridimensionnelle - Google Patents

Procédé de fabrication d'un produit de bio-ingénierie utilisant la bio-impression tridimensionnelle Download PDF

Info

Publication number
WO2019162790A1
WO2019162790A1 PCT/IB2019/051015 IB2019051015W WO2019162790A1 WO 2019162790 A1 WO2019162790 A1 WO 2019162790A1 IB 2019051015 W IB2019051015 W IB 2019051015W WO 2019162790 A1 WO2019162790 A1 WO 2019162790A1
Authority
WO
WIPO (PCT)
Prior art keywords
bioink
scaffold
concentration
ranges
cells
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/IB2019/051015
Other languages
English (en)
Inventor
Alok Medikepura ANIL
Piyush PADMANABHAN
Pooja VENKATESH
Ratandeep Singh BANSAL
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.)
Next Big Innovation Labs Private Ltd
Original Assignee
Next Big Innovation Labs Private Ltd
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 Next Big Innovation Labs Private Ltd filed Critical Next Big Innovation Labs Private Ltd
Publication of WO2019162790A1 publication Critical patent/WO2019162790A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y70/00—Materials specially adapted for additive manufacturing
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y80/00—Products made by additive manufacturing
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction

Definitions

  • the subject described herein in general, relates to a process of three-dimensional (3D) bioprinting. More particularly, but not exclusively, the subject matter relates to a process for 3D bioprinting a tissue and increasing cell viability.
  • 3D bioprinting The process of utilization of 3D printing technology to combine cells with growth factors and biomaterials to develop biomedical parts that imitate natural tissue characteristics is known as three-dimensional (3D) bioprinting.
  • 3D bioprinters also adopt the layer by layer method of depositing bioink to create tissue-like structures, generally referred to as scaffolds.
  • the scaffolds may be further incubated to obtain bioengineered products.
  • the cells within the bioink are subjected to sudden change in temperature, which may harm or kill the cells.
  • the bioink may be relatively highly viscous, thereby requiring relative high pressure to extrude the bioink, which in turn may damage cells within the bioink and adversely impact cell viability.
  • the bioink may be relatively less viscous, due to which the extruded bioink may fail to form a solid structure with desired accuracy or may require excessive use of power to cool the deposited bioink to form a solid structure.
  • a first set of layers of bioink are deposited, followed by crosslinking, and thereafter a second set of layers of bioink are deposited over the first set. It has been observed that, it is challenging to obtain accurate alignment between the first set of layers and the second set of layers. Misalignment has an adverse impact on cell proliferation, and eventually impacts the end product. Further, the instant technique is also found to be relatively time consuming, since there are processing steps to be carried out in between deposition of sets of layers.
  • a method for fabricating a bioengineered product using three-dimensional bioprinting comprising a) preparing a bioink composition, wherein the bioink is prepared by mixing, cell carrier, serum based nutritional supplement and cryoprotectant. The temperature of the prepared bioink is altered to modify the viscosity of the bioink. In the next step the bioink is deposited onto a print plate to form a scaffold and one or more crosslinker is added to the scaffold.
  • FIG. l is a flowchart explaining the 3D bioprinting process, in accordance with an embodiment
  • FIG. 2A illustrates a criss-cross pattern of the scaffold, in accordance with an embodiment
  • FIG. 2B illustrates a lattice pattern of the scaffold, in accordance with an embodiment.
  • the terms“a” or“an” are used, as is common in patent documents, to include one or more than one.
  • the term“or” is used to refer to a non exclusive“or”, such that“A or B” includes“A but not B”,“B but not A”, and“A and B”, unless otherwise indicated.
  • the bioengineered product is developed using living cells.
  • the method includes preparing a bioink composition by mixing the cells, serum based nutritional supplement, cell carrier and cryoprotectant.
  • the temperature of the bioink is altered to modify the viscosity of the bioink.
  • the bioink is deposited onto a print plate to form a scaffold.
  • one or more crosslinker is added to the scaffold to provide structural and chemical integrity to the scaffold.
  • the scaffold is rested to allow crosslinking for a specific period depending on the type of crosslinker used.
  • the scaffold is then washed with another solution to remove residual crosslinker.
  • the scaffold is then incubated for culturing the cells to eventually form the bioengineered product.
  • a method for fabricating bioengineered product using three dimensional bioprinting process.
  • living cells may be added to a solution comprising serum based nutritional supplement and cryoprotectant.
  • the cells are selected based on the bioengineered product or scaffold that has to be fabricated.
  • the cells may be derived from a group consisting of an epithelial, muscular, nervous, or connective tissue, or a suitable combination thereof.
  • the cells are obtained from healthy or diseased donor.
  • the cells may be genetically engineered cells, including induced pluripotent stem cells (iPSCs) or disease specific model cells.
  • iPSCs induced pluripotent stem cells
  • the tissue specific cells may be derived from a tissue selected from a group consisting of liver, gastrointestinal, pancreatic, kidney, lung, tracheal, vascular, skeletal muscle, cardiac, skin, smooth muscle, connective tissue, corneal, genitourinary, breast, reproductive, endothelial, epithelial, fibroblast, neural, Schwann, adipose, bone, bone marrow, pericytes, mesothelial, endocrine, stromal, lymph, and blood.
  • the dilution of the solution is based on the cell count.
  • 1 ml of the solution may be used for a cell count of 1 million.
  • the solution is prepared such that it does not affect the concentration of the cell carrier.
  • the serum based nutritional supplement and cryoprotectant solution provides the cells a protective coating. This helps in protecting the cells from being exposed to sudden decrease in temperature during the bioprinting process. The absence of a protective coating may kill or harm the cells.
  • 1 ml of the bioink comprises about 1 * 10 L 4 -9* 10 L 6 cells.
  • the quantity of cell count may depend on the surface area of the desired scaffold to be printed.
  • 1 ml of the bioink comprises 2 -20 v/v % of serum based nutritional supplement.
  • the serum based nutritional supplement may be biological fluids like serum (2% - 20% v/v).
  • the serum is for example, but not limited to, bovine serum (cow), chicken serum, caprine (goat), equine (horse), human, ovine (sheep), porcine (pig) or rabbit sera.
  • 1 ml of the bioink comprises 0.01- 20 v/v % of chemically defined supplements.
  • the serum-based supplements may be substituted by the chemically defined supplements.
  • the chemically defined supplements may be a tissue extract for example but not limited to bovine pituitary extract (0.1% - 2% v/v) or growth factors or growth hormones or growth regulating factor for example but not limited to EGF (0.05 - 100 ng/ml), VEGF (2 - 50 ng/ml), hydrocortisone (0.1 - 20 pg/ml), insulin (0.5 - 50pg/ml), epinephrine (0.05 - 4pg/ml), transferrin (1 - 25pg/ml), heparin, non-essential amino acids, PDGF (1 - 50 ng/ml), TGF (0.001 - 20pg/ml).
  • EGF 0.05 - 100 ng/ml
  • VEGF 2 - 50 ng/ml
  • hydrocortisone 0.1 - 20 pg/ml
  • insulin 0.5 - 50pg/ml
  • epinephrine 0.05 - 4pg
  • 1 ml of the bioink comprises 2% -10% v/v of cryoprotectant.
  • a single cryoprotectant or a combination of various cryoprotectants may be used to prepare the bioink.
  • the cryoprotectant may be Dimethyl Sulfoxide DMSO (0.5 % - 10% v/v), glycerol (1% - 5% v/v), hydroxyethyl starch (0.1 - 10%), PEG or combinations thereof.
  • the concentration of the various cryoprotectants when used in combination to form the cryoprotectant having a minimum concentration of 2% v/v are for example, but not limited to, a) DMSO 0.5% and glycerol 1.5%; b) Glycerol 1% and DMSO 1%; c) DMSO 0.5%, hydroxy ethyl starch 0.1% and glycerol 1.4%.
  • the concentration of the DMSO or any other cryoprotectant is at least 2% v/v, but when used in combination the range may vary as provided above in the form of examples.
  • the cryoprotectant having the maximum concentration i.e. 10% v/v may be prepared by using various combination of the cryoprotectants. When a single cryoprotectant is used to prepare the bioink, the concentration of the single cryoprotectant is maximum 10% v/v.
  • 1 ml of the bioink comprises 1-12 w/v % of cell carrier.
  • the cell carrier is prepared at step 104.
  • the cell carrier is prepared by dissolving biomaterials in a solvent.
  • the cell carrier is prepared by dissolving one or more biomaterials in a solvent.
  • the solvent may be a combination of water (0% -75% v/v) and culture media (100% -25% v/v), for example but not limited to, 10% water and 90% media.
  • water may be replaced with PBS, HBSS, TBS, HEPES or MOPS, and the media with DMEM.
  • the solvent may be prepared using the following components NaOH (0.8%), HC1 (1.8%), KOH (0.3%), Nicotinic acid (5mg/L), S02 (1%), Ethanol (2%), Chloroform (1%), Dichloromethane (1%), Carbon tetrachloride (2.5mM), Benzene (l5mM), Toluene (lOmM), Ethylbenzene (lOmM), Xylene (5mM), Acetone (5%), Dimethyl formamide (0.5%), Glycerol (5%), Polyethelyene glycol i.e, PEG (2%) , triethylene glycol (1.5%) or combination thereof.
  • the range disclosed for each of the component is the maximum range.
  • the above listed components may be used in combination with media and water (water could be replaced PBS, HBSS, TBS, HEPES or MOPS) or with media alone.
  • the combinations are for example, but not limited to, a) PEG (2%) and media 98%; b) PEG (2%), water (25%) and media (73%); c) PEG (1%), ethanol (1%), water (24%) and media (73%); d) Ethanol (1%) and media (99%); and e) Acetone (2%) and DMEM (98%).
  • the selection of the culture media may depend on the cell type used.
  • the culture media could be for example, but not limited to, Dulbecco's Modified Eagle's Medium (DMEM), Roswell Park Memorial Institute medium (RPMI), Minimum Essential Media (MEM), Iscove's Modified Dulbecco's Medium (IMDM), Reduced-Serum Medium (Opti - MEM), Minimum Essential Media alpha (a MEM), Me Coy’s 5 A and modifications of these media.
  • the biomaterial may be natural polymers, synthetic polymers or a combination of both natural and synthetic polymers.
  • the biomaterial may be a naturally occurring polymer selected from the group consisting of collagen, fibrin, chitosan, alginate, oxidized alginate, starch, hyaluronic acid, laminin, silk fibroin, agarose, gelatine, glycans, and combinations thereof.
  • the biomaterial when used in combination the concentration of each of the biomaterials may vary based on the type of the biomaterial used.
  • a list of various natural biomaterial with the concentration range is provided when the cell carrier is prepared using one or more biomaterials in combination, such as, but not limited to, collagen (0.1- 5% w/v), gelatin (1-12 % w/v), agarose (0.2-2 % w/v), agar (0.01- 1 % w/v), alginate (0.05- 2% w/v), hyaluronic acid (0.01- 0.5% w/v), fibrin (0.5-l0%)or combinations thereof.
  • the biomaterial may be a synthetic polymer selected from the group consisting of polyphosphazene, polyacrylic acid, polymethacrylic acid, polylactic acid (PLA), polyglycolic acid (PGA), poly (lactic-co-glycolic acid) (PLGA), polyorthoester (POE), polycaprolactone (PCL), polyamine acid (such as polylysine), degradable polyurethane, copolymers, and combinations thereof.
  • PPA polylactic acid
  • PGA polyglycolic acid
  • PLGA poly (lactic-co-glycolic acid)
  • POE polyorthoester
  • PCL polycaprolactone
  • PCL polyamine acid (such as polylysine), degradable polyurethane, copolymers, and combinations thereof.
  • the choice of the solvent may depend on the polymer used.
  • the biomaterials are mixed in the solvent using a shaker for about 5 minutes to 60 minutes, and for about 1 hour to 24 hours for natural polymers and synthetic polymers respectively.
  • the study reveals that the temperature of the cell carrier is maintained between 12 degree Celsius 27.9 degree Celsius to obtain the desired viscosity and the size of the needle gauge used in the bioprinter ranges from 20G-30G.
  • the experimental study performed with different concentration of the cell carrier reveals that lower the concentration of the cell carrier, lower is the temperature to be maintained to obtain the desired viscosity.
  • the cells that were immersed in serum based nutritional supplement and cryoprotectant solution
  • the quantity of cell mixture may be negligible compared to the quantity of cell carrier.
  • the bioink may be prepared by mixing the living cells, the cell carrier, the serum based nutritional supplement and the cryoprotectant.
  • the bioink is processed, for example by regulating or altering the temperature of the bioink that correspond to a range of preferred viscosity. In an embodiment, the temperature of the bioink is altered by cooling.
  • the bioink is immersed in ice prior to the bioprinting process for a predetermined time. In an embodiment, the predetermined time may be three minutes to four minutes. In another embodiment, the predetermined time varies accordingly depending on the concentration of the cell carrier used for the preparation of the bioink (Refer table 1 and 2). For example, the bioink is immersed for about half an hour if the concentration of the cell carrier is 3% w/v and for about 30 seconds if the concentration of the cell carrier is 12% w/v.
  • bioink semi gel like The immersion of bioink in the ice makes the bioink semi gel like.
  • This semi gel composition gives the bioink the desired viscosity that is required for bioprinting process. If the viscosity is not right, then either the bioink may be too thick or too watery. High viscosity of the bioink may result in applying excessive force to extrude the bioink. Excessive force may lead to high shear within the bioink which may end up harming or killing the cells. On the other hand, if the viscosity of the bioink is too less, the scaffold eventually formed may not be structurally stable and may lack accuracy. Thus, keeping the bioink in ice for a predetermined time just before adding it to the extruder, provides the bioink with stable characteristics.
  • the bioink may be directly placed in the extruder to cool the bioink.
  • the extruder may comprise a cooling jacket, wherein the cooling jacket may provide the extruder with the required cooling. This may keep the bioink within the extruder semi gel like, to provide the bioink the desired viscosity (refer table 1 and table 2).
  • the inner diameter of the extruder needle (of the extruder) may be varied to extrude bioinks of different viscosities.
  • the inner diameter of the extruder needle may be reduced to extrude the bioink with lower viscosity.
  • the inner diameter of the extruder needle may be increased to extrude the bioink with higher viscosity.
  • the bioink (that was kept in the ice for 3-4 minutes) is introduced into the extruder of the 3D bioprinter.
  • the print plate prior to extruding, the print plate is allowed to cool.
  • the print plate is cooled to about -10 degree Celsius to -25 degree Celsius.
  • the print plate may be cooled via cooling a mechanism for example but not limited to a peltier device.
  • the print plate may be a petri plate, a quartz plate or a glass slide.
  • the print plate may be made of metals for example but not limited to aluminium and stainless steel. Further, the print plate should be sterile, non- corrosive having a smooth flat surface.
  • the scaffold may be printed in any of the patterns that exists or that may exist in the future.
  • the scaffold may be printed in a lattice pattern or a criss-cross pattern or a combination of both lattice pattern and criss-cross pattern. Referring to FIGs.2A-2B illustrates the criss-cross pattern and lattice pattern of the scaffold.
  • the criss-cross pattern of the scaffold increases contact between the cells. Further, the criss cross pattern also decreases the cell migration distance between the pores.
  • the extruder may extrude the bioink using any one of the existing extruder mechanisms such as, pneumatic, hydraulic, mechanical, electronic, combination of the existing mechanism or the mechanisms that may be made available in future.
  • the scaffold is allowed to rest on the print plate (without switching off the peltier) for a predetermined time.
  • the predetermined time may be 2-3 minutes.
  • the crosslinker is added to the scaffold.
  • the crosslinker is added to provide structural cohesion to the scaffold.
  • the crosslinker may be added either manually or mechanically. Manual crosslinking may be done using a pipette. Mechanical or automated crosslinking may be done using a mechanism that may be similar to the extruder mechanism.
  • the scaffold is allowed to rest on the print plate (without switching off the peltier) for a predetermined time. In an embodiment, the predetermined time may be 5-6 minutes.
  • the bioprinted scaffold may undergo physical crosslinking, chemical crosslinking, photo crosslinking or combinations thereof.
  • the chemical cross linking may be performed by any cationic or anionic or non-ionic cross-linkers.
  • the scaffold may be crosslinked by calcium, magnesium, sodium, chloride, alginate, or any combination thereof.
  • enzymatic cross-linkers may be used for cross linking the bioprinted scaffold.
  • the scaffold after allowing to be rested on the print plate for the predetermined time, is stored at a predetermined temperature for a predetermined time.
  • the predetermined temperature may range from -10 degree Celsius to -196 degree Celsius.
  • the predetermined temperature may be in the range of -10 degree Celsius to -40 degree Celsius and the predetermined time may be 6 to 18 hours.
  • the predetermined time for which the scaffold is kept for crosslinking may depend on the quantity and concentration of the crosslinker and the cell carrier.
  • the scaffold is washed with a solution to remove any residual crosslinker.
  • the solution may be Phosphate Buffered Saline (PBS).
  • the scaffold is dipped in a media and introduced into an incubator that is maintained at a predetermined temperature. Further, the amount of C0 2 within the incubator may be maintained at a predetermined level for culturing the cells.
  • the predetermined temperature may range from 37 degree Celsius to 39 degree Celsius and the predetermined level of C0 2 maintained may range from 5% to 7% v/v. The tissue thus obtained may have high cell viability.
  • the skin tissue may include keratinocytes, melanocytes and fibroblasts.
  • skin tissue comprising of only keratinocytes may be 3D bio printed or skin tissue comprising of only fibroblasts may be 3D bio printed or skin tissue comprising of a combination of both fibroblasts and keratinocytes may be 3D bio printed.
  • Skin cells are prepared and mixed with the solution comprising the serum based nutritional supplement and cryoprotectant. to protect the skin cells from being exposed to sudden decrease in temperature. Meanwhile, the cell carrier is prepared by dissolving the biomaterial in a combination of water and media.
  • the cells that were immersed in the solution comprising the serum based nutritional supplement and cryoprotectant.
  • the mixture of cells and cell carrier may be called as bioink.
  • the bioink is immersed in ice for 3-4 minutes and placed in the extruder.
  • the peltier device is turned on before printing the bioink so that the surface of the print plate is cooled enough.
  • the bioink is then deposited in layers. The number of layers of the bioink to be deposited may depend on the nature of the tissue that may be developed.
  • the scaffold is left on the print plate for about 2-3 minutes, with the print plate kept on.
  • the crosslinker is then added to the scaffold for crosslinking.
  • Crosslinking helps in giving structural cohesion to the tissue.
  • the scaffold is left on the print plate for 5-6 minutes, with the print plate still cold.
  • the print plate is switched off and the scaffold is removed from the print plate, sealed and stored at 4°C to -20°C for about 6 hours 12 hours.
  • the scaffold is then immediately washed with PBS to remove any residual crosslinkers if there are any.
  • the scaffold is then dipped in the media and sent to the incubator for culturing the skin cells.
  • the incubator is maintained at 37°C and 5% C0 2 for the duration of the culture.
  • the volume of new media added is such that the tissue is brought to Air Liquid Interface (ALI), i.e., so that keratinocytes can proliferate and differentiate.
  • ALI Air Liquid Interface
  • the media is changed for every 2-3 days for replenishing the required nutrients.
  • the scaffold is kept in the incubator until the tissue is formed.
  • the skin tissue thus obtained has high cell viability.
  • the present invention overcomes the drawbacks of the conventional bioprinting processes, by providing an improved way of bioprinting to achieve high cell viability.
  • the present invention as discussed in this document with respect to different embodiments will be advantageous at least in protecting the cells from being exposed to sudden change in the printing parameters such as temperature and pressure during the bioprinting process. This may prevent the killing or harming of the cells during the bioprinting process.
  • the present invention is advantageous in providing the bioink with the required viscosity. This helps in providing the bioink the required stability during the bioprinting process. Also, the present invention is advantageous in bioprinting multiple layers in a single extrusion process, thus preventing multiple manual extrusion processes.
  • the present invention reduces the overall cost of developing the tissue, reduces the amount of materials required, increases the precision during the bioprinting process, provides proper spatial location of the cells. Additional advantages not listed may be understood by a person skilled in the art in light of the embodiments disclosed above.

Landscapes

  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biomedical Technology (AREA)
  • Materials Engineering (AREA)
  • Manufacturing & Machinery (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Dermatology (AREA)
  • Botany (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Transplantation (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Zoology (AREA)
  • Cell Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Materials For Medical Uses (AREA)

Abstract

La présente invention concerne la préparation de bio-encre, la bio-encre étant préparée par mélange de cellules, de support de cellules, de supplément à base de sérum, et d'agent cryoprotecteur. La température de la bio-encre est altérée pour modifier la viscosité de la bio-encre. La bio-encre ayant la viscosité souhaitée est déposée sur une plaque d'impression pour former une charpente. Dans l'étape suivante, un ou plusieurs agents de réticulation sont ajoutés à la charpente obtenue ci-dessus. L'agent de réticulation est ajouté pour fournir l'intégrité structurelle et chimique à la charpente. La charpente est laissée au repos pour permettre la réticulation sur une période spécifique. La charpente est ensuite lavée avec une solution saline tamponnée au phosphate (PBS) pour retirer l'agent de réticulation résiduel. La charpente est ensuite incubée pour cultiver les cellules afin de former éventuellement le produit de bio-ingénierie.
PCT/IB2019/051015 2018-02-21 2019-02-08 Procédé de fabrication d'un produit de bio-ingénierie utilisant la bio-impression tridimensionnelle Ceased WO2019162790A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IN201841006622 2018-02-21
IN201841006622 2018-02-21

Publications (1)

Publication Number Publication Date
WO2019162790A1 true WO2019162790A1 (fr) 2019-08-29

Family

ID=67687009

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB2019/051015 Ceased WO2019162790A1 (fr) 2018-02-21 2019-02-08 Procédé de fabrication d'un produit de bio-ingénierie utilisant la bio-impression tridimensionnelle

Country Status (1)

Country Link
WO (1) WO2019162790A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022261534A3 (fr) * 2021-06-11 2023-01-19 University Of Maryland, Baltimore Échafaudages par bioencre chargés de cellules tridimensionnelles et leurs procédés de production dans des conditions cryogéniques pour l'ingénierie tissulaire

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170022540A1 (en) * 2015-07-24 2017-01-26 University Of South Florida Bioink for three-dimensional biomaterial printing
WO2017023865A1 (fr) * 2015-07-31 2017-02-09 Techshot, Inc. Système de biofabrication, procédé, et matériel de bioimpression 3d dans un environnement à gravité réduite
US20170130192A1 (en) * 2015-11-09 2017-05-11 Organovo, Inc. Methods for tissue fabrication

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170022540A1 (en) * 2015-07-24 2017-01-26 University Of South Florida Bioink for three-dimensional biomaterial printing
WO2017023865A1 (fr) * 2015-07-31 2017-02-09 Techshot, Inc. Système de biofabrication, procédé, et matériel de bioimpression 3d dans un environnement à gravité réduite
US20170130192A1 (en) * 2015-11-09 2017-05-11 Organovo, Inc. Methods for tissue fabrication

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022261534A3 (fr) * 2021-06-11 2023-01-19 University Of Maryland, Baltimore Échafaudages par bioencre chargés de cellules tridimensionnelles et leurs procédés de production dans des conditions cryogéniques pour l'ingénierie tissulaire

Similar Documents

Publication Publication Date Title
Kozlowski et al. Towards organoid culture without Matrigel
Choudhury et al. Organ-derived decellularized extracellular matrix: a game changer for bioink manufacturing?
Liu et al. Porous nanofibrous poly (L-lactic acid) scaffolds supporting cardiovascular progenitor cells for cardiac tissue engineering
Lavik et al. Fabrication of degradable polymer scaffolds to direct the integration and differentiation of retinal progenitors
Bellas et al. In vitro 3D full‐thickness skin‐equivalent tissue model using silk and collagen biomaterials
Biazar et al. Chitosan–cross-linked nanofibrous PHBV nerve guide for rat sciatic nerve regeneration across a defect bridge
Lode et al. Additive manufacturing of collagen scaffolds by three-dimensional plotting of highly viscous dispersions
US11471285B2 (en) Tissue grafts and methods of making and using the same
JP7007267B2 (ja) 組織製造のための改善された方法
US20110052693A1 (en) Method for producing artificial skin
Ladd et al. Development of intestinal scaffolds that mimic native mammalian intestinal tissue
EP3004331A1 (fr) Utilisation de cellules mésothéliales en bio-ingénierie tissulaire et de tissus artificiels
Senturk et al. 3D bioprinting of tyramine modified hydrogels under visible light for osteochondral interface
CN112823030A (zh) 动脉内皮细胞种植的血管移植物的生成
Hu et al. Patient-specific cardiovascular progenitor cells derived from integration-free induced pluripotent stem cells for vascular tissue regeneration
US20200289702A1 (en) Microspheres Containing Decellularized Donor Tissue and Their Use in Fabricating Polymeric Structures
Worthington et al. Differentiation of induced pluripotent stem cells to neural retinal precursor cells on porous poly-lactic-co-glycolic acid scaffolds
Zand et al. Successful 3D culture and transplantation of mouse isolated preantral follicles in hydrogel of bioengineered Wharton’s jelly
WO2019162790A1 (fr) Procédé de fabrication d'un produit de bio-ingénierie utilisant la bio-impression tridimensionnelle
CN102631710A (zh) 多通道多层细胞结构的复合组织器官前体的制备方法
US20220364049A1 (en) Composition and methods for culturing retinal progenitor cells
EP4331673A1 (fr) Agrégat sous forme de cordon de cellules de l'épithélium pigmentaire rétinien, dispositif ainsi que procédé de fabrication de celui-ci, et remède comprenant cet agrégat sous forme de cordon
CN110607278A (zh) 肿瘤细胞培养物及其应用
KR20200012723A (ko) 광가교제 없이 3d 프린팅할 수 있는 세리신 기반 바이오잉크 조성물
Wismayer et al. Improving cellular migration in tissue-engineered laryngeal scaffolds

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: 19756851

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: 19756851

Country of ref document: EP

Kind code of ref document: A1