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WO2012023093A1 - Echafaudage polymère de polycaprolactone tridimensionnel - Google Patents

Echafaudage polymère de polycaprolactone tridimensionnel Download PDF

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Publication number
WO2012023093A1
WO2012023093A1 PCT/IB2011/053586 IB2011053586W WO2012023093A1 WO 2012023093 A1 WO2012023093 A1 WO 2012023093A1 IB 2011053586 W IB2011053586 W IB 2011053586W WO 2012023093 A1 WO2012023093 A1 WO 2012023093A1
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human
cells
multiple myeloma
scaffold
primary
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Inventor
Pierfrancesco Tassone
Filippo Causa
Pierosandro Tagliaferri
Teresa Calimeri
Maria Teresa Di Martino
Edmondo Battista
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Fondazione T Campanella - Coe
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Fondazione T Campanella - Coe
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/027—New or modified breeds of vertebrates
    • A01K67/0271—Chimeric vertebrates, e.g. comprising exogenous cells
    • 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
    • A61L27/14—Macromolecular materials
    • A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
    • 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
    • A61L27/3604—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 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • 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
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/56—Porous materials, e.g. foams or sponges
    • 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
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0693—Tumour cells; Cancer cells
    • C12N5/0694—Cells of blood, e.g. leukemia cells, myeloma cells
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2227/00—Animals characterised by species
    • A01K2227/10—Mammal
    • A01K2227/105—Murine
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00—Animals characterised by purpose
    • A01K2267/03—Animal model, e.g. for test or diseases
    • A01K2267/0331—Animal model for proliferative diseases
    • 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
    • C12N2502/00—Coculture with; Conditioned medium produced by
    • C12N2502/13—Coculture with; Conditioned medium produced by connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
    • C12N2502/1394—Bone marrow stromal cells; whole marrow
    • 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
    • C12N2503/00—Use of cells in diagnostics
    • C12N2503/04—Screening or testing on artificial tissues
    • 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
    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/30—Synthetic polymers
    • C12N2533/40—Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers

Definitions

  • the present invention refers to chemistry field and to its applications in medicine.
  • the present invention refers to a three-dimensional (3-D) poly- ⁇ - caprolactone polymeric (PCL) scaffold, to its use for in vivo engraftment of human multiple myeloma (MM) cells within an allogeneic as well as autologus human bone marrow microenvironment (HuBMM).
  • the present invention also refers to a non-human animal, preferably a mouse, carrying said scaffold.
  • MM cells home to the BM (bone marrow) and adhere to ECM (extracellular matrix) proteins and to BMSCs (Bone Marrow Stromal Cells)
  • BMSCs Blood Cells 1992 Nov 1 ; 80(9): 2306-2314; Okado T, Hawley RG.
  • the close cross-talk of MM cells with the non-tumor compartment within the bone marrow plays a key role in supporting tumor growth, survival and development of drug resistance (Ribatti D, Nico B, Vacca A. Importance of the bone marrow microenvironment in inducing the angiogenic response in multiple myeloma. Oncogene 2006 Jul 20; 25(31 ): 4257- l 4266; Mitsiades CS, Mitsiades NS, Munshi NC, Richardson PG, Anderson KC.
  • the role of the bone microenvironment in the pathophysiology and therapeutic management of multiple myeloma interplay of growth factors, their receptors and stromal interactions.
  • the SCID-hu model which is based on the implantation of a human fetal bone chip in a SCID mouse, has been the most biologically relevant system since it reproduces in vivo a HuBMM for the successful engraftment and growth of human primary MM cells (Urashima M, Chen BP, Chen S, Pinkus GS, Bronson RT, Dedera DA, ei al.
  • the SCID-hu system is still one of the most relevant preclinical model of human disease.
  • This model offers several advantages over conventional subcutaneous (LeBlanc R, Catley LP, Hideshima T, Lentzsch S, Mitsiades CS, Mitsiades N, et al.
  • Proteasome inhibitor PS-341 inhibits human myeloma cell growth in vivo and prolongs survival in a murine model. Cancer Res 2002 Sep 1 ; 62(17): 4996-5000; Tassone P, Gozzini A, Goldmacher V, Shammas MA, Whiteman KR, Carrasco DR, et al.
  • PCLS three-dimensional (3-D) ⁇ - ⁇ -caprolactone polymeric scaffold
  • Object of the present invention is a three-dimensional scaffold consisting of poly- ⁇ - caprolactone and its use for in vivo engraftment of human multiple myeloma (MM) cells in a non human animal, in particular a mouse. Therefore, the present invention provides a novel in vivo human MM model based upon the implantation into a recipient non-human animal of said three-dimensional (3-D) bone-like ⁇ - ⁇ -caprolactone polymeric scaffold (PCLS) which then allows for in vivo engraftment of human MM cells in a HuBMM.
  • PCLS three-dimensional scaffold
  • the present invention provides a three-dimensional (3-D) ⁇ - ⁇ -caprolactone polymeric scaffold (PCLS) which can be used for in vivo engraftment of human MM cells in an allogeneic, or preferably, an autologous HuBMM.
  • PCLS three-dimensional ⁇ - ⁇ -caprolactone polymeric scaffold
  • Different scaffolds and methods to obtain polymeric scaffolds are already known (WO2005/020849, WO2008/041854, WO2009/054006, WO2010/067086).
  • Object of the present invention is a three-dimensional scaffold consisting of poly- ⁇ - caprolactone, optionally pre-coated with human bone marrow stromal cells (BMSCs) isolated from a multiple myeloma patient.
  • BMSCs bone marrow stromal cells
  • Another object of the present invention is the use of the above scaffold for engraftment and in vivo growth of human primary or IL-6-dependent multiple myeloma cells in a non-human animal, preferably a mouse.
  • the scaffold is pre-coated with human BMSCs, and said MM cells are immunoselected CD138 + primary multiple myeloma cells isolated from patient or established IL-6/bone marrow (BM) dependent INA-6 multiple myeloma cells.
  • BM bone marrow
  • the scaffold is not pre-coated with human BMSCs, and said primary MM cells are comprised in human bone marrow mononuclear cells (BMMCs) isolated from patient.
  • BMMCs bone marrow mononuclear cells
  • a further object is a method for the growth of human primary or IL-6-dependent multiple myeloma cells comprising the steps of: a) seeding human bone marrow stromal cells isolated from a multiple myeloma patient in the empty scaffold; b) engrafting primary multiple myeloma cells isolated from a patient or IL-6- dependent multiple myeloma cells within said scaffold in a non-human animal, preferably a mouse.
  • step a) is preferably performed in vitro.
  • Another object of the present inventions is a method for the growth of human primary or IL-6-dependent multiple myeloma cells comprising the steps of: a) seeding human bone marrow stromal cells isolated from a multiple myeloma patient in the empty scaffold; b) engrafting immunoselected CD138 + primary multiple myeloma cells isolated from a patient or IL-6/BM dependent INA-6 multiple myeloma cells within said scaffold in a non-human animal, preferably a mouse.
  • step a) is preferably performed in vitro.
  • a further object of the invention is a method for the growth of human primary multiple myeloma cells comprising the step of seeding human bone marrow mononuclear cells isolated from patient, which comprise primary multiple myeloma cells, in the scaffold not pre-coated with human BMSCs, in a non-human animal, preferably a mouse.
  • the above step of seeding human bone marrow mononuclear cells is preferably performed in vitro.
  • Said human bone marrow mononuclear cells (BMMCs) isolated from patient comprise primary MM cells and BMSCs.
  • Another object is a method for the growth of human primary or IL-6-dependent multiple myeloma cells comprising the step of engrafting immunoselected CD138 + primary multiple myeloma cells isolated from a patient or IL-6/BM dependent INA-6 multiple myeloma cells within the pre-coated scaffold, in a non-human animal, preferably a mouse.
  • An object of the invention is also a non-human animal, preferably a mouse, carrying the scaffold of the invention, optionally pre-coated with human bone marrow stromal cells (BMSCs) isolated from a multiple myeloma patient or in vitro seeded with bone marrow mononuclear cells (BMMCs), containing both primary MM cells and their autologous BMSCs, isolated from MM patient.
  • BMSCs bone marrow stromal cells
  • a preferred mouse is the SCID-synth-hu.
  • the MM-bearing SCID-synth-hu has herein shown to be a suitable model for large scale drug screening.
  • a further important observation is the detection of angiogenic events in retrieved scaffolds.
  • the present inventors observed vessels of variable size in scaffolds precoated with BMSCs as well as in scaffolds seeded with BMMCs and the endothelial cells reacted with a specific anti-human CD31 mAb.
  • the findings of the present inventors support the use of the SCID-synth-hu as first and unique system for in vivo engraftment of human primary MM cells within their allogeneic or, preferably, autologous adult HuBMM.
  • the system of the present invention provides significant advantages including: i) unlimited availability of PCLSs; ii) the potential for dissecting the biological events within the HuBMM since the 3-D recipient can be precoated with selected or genetically manipulated BM cell populations; Hi) the possibility to engraft primary MM cells in a non-fetal adult autologous HuBMM.
  • the present model represents the first system for in vivo expansion of human MM cells on a synthetic platform.
  • the SCID-synth-hu is therefore a unique tool for large scale in vivo preclinical evaluation of novel agents targeting MM in its autologous HuBMM and a novel resource for translational research in the experimental treatment of this still incurable disease.
  • Object of the present invention is a three-dimensional scaffold consisting of poly- ⁇ - caprolactone, optionally pre-coated with human bone marrow stromal cells (BMSCs) isolated from a multiple myeloma patient.
  • BMSCs bone marrow stromal cells
  • IL-6-dependent multiple myeloma cells are used to confirm the goodness of reproduction of the microenvironment. This cell line has not completely lost the dependence from the microenvironment. In fact these cells grow in vitro only in presence of BMSCs or human IL-6 and they don't grow if subcutaneously or endovenously injected.
  • the scaffold is pre-coated with human BMSCs, and said MM cells are immunoselected CD138 + primary multiple myeloma cells isolated from patient or established IL-6/bone marrow (BM) dependent INA-6 multiple myeloma cells.
  • BM bone marrow
  • a "pre-coated scaffold” is a three-dimensional scaffold consisting of ⁇ - ⁇ -caprolactone pre-coated with human bone marrow stromal cells (BMSCs) isolated from a multiple myeloma patient.
  • BMSCs bone marrow stromal cells
  • an "empty scaffold” is the three-dimensional scaffold consisting of poly-£-caprolactone.
  • BMSCs and said MM cells are isolated from different subjects in order to avoid ex-vivo manipulation of primary cells (allogeneic HuBMM).
  • the scaffold is not pre-coated with human BMSCs, and said primary MM cells are comprised in human bone marrow mononuclear cells (BMMCs), isolated from patient, comprising primary MM cells and BMSCs.
  • BMMCs human bone marrow mononuclear cells
  • the HuBMM can be defined as autologous. All the above uses are in a non-human animal, preferably a mouse
  • NaCI particles were sieved into a specific size range (212-300 mm) and added to form a homogeneous mix (weight ratio of 9/1 ). The mixture was then poured into a silicon pipe with an inner diameter of 3 mm and dipped in ethanol (J.T. Baker, 99.9%) to allow solvent extraction. Further immersion in bidistilled water (10 days) was performed to allow sodium chloride crystal dissolution (salt leaching) and to remove residual traces of solvents. Finally, after drying, samples were pulled out from the mould and cut into 3 x 7 mm cylinders. The pore size was evaluated by an image analysis procedure and porosity degree though gravimetric methods.
  • micrographs obtained by Scanning Electron Microscopy (SEM) of cross-section surfaces were processed by an imaging software (ImageJ 1.35).
  • images require the conversion from analogical to digital image: the 8-bit conversion allows obtaining a monochromatic image with 256 grey levels where the level 0 corresponds to pure black and the highest level 255 to pure white.
  • the adequate definition of the threshold level allows to define the optimal contrast and brightness to calculate the pore features.
  • Means and standard deviations of pore volume fraction and pore size have been determined from 10 measurements per micrograph.
  • the density of the porous scaffold was calculated by measuring the dimensions and the mass of the scaffold (gravimetric method) and was calculated as follows: where m is the mass and V is the volume of the porous structure.
  • OP9 mouse stromal cells were kindly provided by Dr. Giovanni Morrone (University of Magna Graecia, Catanzaro, Italy) and cultured in a-MEM (GIBCO) containing 10% heat inactivated fetal bovine serum (FBS), 50 g/ml vitamin C and 0,07% ⁇ -mercaptoethanol.
  • FBS heat inactivated fetal bovine serum
  • the IL-6/BM dependent INA-6 MM cell line was kindly provided by Dr.
  • Renate Burger (University of Kiel, Kiel, Germany) and cultured in the presence of 2,5 ng/ml of human recombinant IL-6 (Sigma-Aldrich) as described (Neri P, Tagliaferri P, Di Martino MT, Calimeri T, Amodio N, Bulotta A, ei al.
  • IL-6 human recombinant IL-6
  • Combination therapy with interleukin-6 receptor superantagonist Sant7 and dexamethasone induces antitumor effects in a novel SCID-hu In vivo model of human multiple myeloma.
  • the IL-6 receptor super- antagonist Sant7 enhances antiproliferative and apoptotic effects induced by dexamethasone and zoledronic acid on multiple myeloma cells. Int J Oncol 2002 Oct; 21 (4): 867-873).
  • Human BMSCs were collected from bone borrow aspirates of MM patients after informed consent and cultured in RPMI 1640 (Sigma-Aldrich) with l-glutamine and NaHCO3, containing 20% heat inactivated fetal bovine serum (FBS), 100U/ml_ Penicillin, 1 ⁇ g/ml Streptomycin and 1 ⁇ g/mL Amphotericin B (Sigma-Aldrich). Two thirds of medium were replaced once a week and, after 28 days of culture, cells adherent to culture plastic flasks were considered as BMSCs and were harvested with trypsin.
  • FBS heat inactivated fetal bovine serum
  • Human primary isolated MM cells were obtained from bone marrow aspirates of 8 MM patients and from peripheral blood samples from 2 plasma cell leukemia patients as previously described (Tassone P, Gozzini A, Goldmacher V, Shammas MA, Whiteman KR, Carrasco DR, et al.
  • CD138 + MM cells were selected by immunomagnetic separation (Miltenyi Biotec). Purity of CD138 + MM cells (more than 95%) was assessed by flow cytometry (FACS). Unselected populations of bone marrow mononuclear cells (BMMCs), obtained by Ficoll gradient separation of whole bone marrow aspirate, have been used as alternative sources of primary MM cells and autologous BMSCs.
  • BMMCs bone marrow mononuclear cells
  • CD138+ immunoselected primary MM cells or IL-6/BM dependent INA-6 MM cells were directly injected into the scaffold in vivo in 100 ⁇ of growth medium, 2-3 weeks after the implant of PCLSs which have been previously coated with non autologous BMSCs.
  • BMMCs obtained by Ficoll gradient separation and containing primary unselected CD138+ and their autologous BMSCs, were seeded in vitro into PCLSs which, after 24 h of incubation, were implanted in SCID mouse flank.
  • Mice CB-17 SCID mice (6-8 week-old male) were purchased from Charles River Laboratories and maintained in our Animal Research Facility. Animal studies were approved by our Institutional Veterinary Service and Ethical Committee. Procedures for surgical implantation of PCLSs in mice were similar to those already described for bone chips in SCID-hu mice (Urashima M, Chen BP, Chen S, Pinkus GS, Bronson RT, Dedera DA, ei al.
  • MM engraftment and growth was monitored by paraprotein detection in mouse sera by enzyme-linked immunosorbent assay (ELISA; R&D Systems, Minneapolis, MN), as previously described (Neri P, Tassone P, Shammas M, Yasui H, Schipani E, Batchu RB, ei al. Biological pathways and in vivo antitumor activity induced by Atiprimod in myeloma. Leukemia 2007 Dec; 21 (12): 2519-2526; Tassone P, Goldmacher VS, Neri P, Gozzini A, Shammas MA, Whiteman KR, ei al. Cytotoxic activity of the maytansinoid immunoconjugate B-B4-DM1 against CD138+ multiple myeloma cells. Blood 2004 Dec 1 ; 104(12): 3688-3696).
  • ELISA enzyme-linked immunosorbent assay
  • mice blood was collected from tail vein every two weeks.
  • SCID-synth-hu mice bearing MM were intraperitoneally (i.p.) treated with bortezomib (on days 1 , 4, 8, and 1 1 ) plus dexamethasone (on days 1 to 4), and MM response was monitored by measuring paraprotein levels in mice sera.
  • mAbs including anti-CD138/MI 15 (Dako), anti-CD56/123C3 (Zymed), anti-kappa (Dako), anti-lambda (Dako), anti-CD31/JC70A (Dako), anti- CD34/QBEnd-10 (Dako) and anti-FVIII/E980-1 (Novocastra) were used according to manifacturer's instructions.
  • BMSCs The adherence of BMSCs into PCLS was also evaluated by CLSM. After 21 days of culture in flasks, the cells harvested with trypsin were stained by adding 500 ⁇ L 4,6-diamidino-2-phenylindole (DAPI; 1 : 1000) for 30 minutes at 4°C and then seeded into scaffolds. After 10 days scaffolds were isolated from mice, reduced with the cryostat in slices of 4 ⁇ and then fixed in formaldehyde.
  • DAPI 4,6-diamidino-2-phenylindole
  • the images were digitalized into 512x512 lines and conveyed to a computer for storage and further analysis.
  • PCLS demonstrated interconnected large (100 to 300 ⁇ ) and small pores (1 to 10 ⁇ ). Surface morphology was studied by a Leica Cambridge (Stereoscan S440) scanning electron microscope (Cambridge, UK) at an accelerating potential of 20 Kv. Comparative SEM analysis between a synthetic PCLS and a human femur adult bone chip obtained from orthopedic surgery showed similar microarchitecture.
  • OP9 a mouse BM stromal cell line, to set up the experimental loading conditions. Since OP9 cells efficiently adhered to 3-D surfaces, we next injected PCLSs with human BMSCs derived from MM patients.
  • BMSCs obtained from two different patients adhered and diffused among 3-D interconnected surfaces, starting one week after cell injection, as demonstrated by SEM, H&E staining and confocal laser scanning microscopy. Based on these achievements, it was next evaluated the suitability of these BMSCs-coated PCLSs for engraftment of primary MM cells or of the IL-6/BM dependent INA-6 MM cell line.
  • CD138+ immunoselected primary MM cells from BM aspirates of patients with newly diagnosed or relapsed disease and from peripheral blood of plasma cell leukemia patients or INA-6 cells were immediately injected in vivo in PCLSs previously coated ex-vivo with BMSCs and then implanted in mice.
  • the present invention finds application in the medical field providing a novel in vivo preclinical model of human multiple myeloma (SCIDsynth-hu model) for identification of novel targets and validation of therapeutical agents targeting MM within its autolougous adult milieu.
  • SCIDsynth-hu model novel in vivo preclinical model of human multiple myeloma

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Abstract

La présente invention a trait à un échafaudage tridimensionnel qui est constitué de poly-ε-caprolactone, éventuellement préenrobé avec des cellules stromales de moelle osseuse humaine, afin d'être utilisé pour une prise de greffe et une croissance in vivo de cellules de myélome multiple dépendant d'IL6 ou primaire humain chez un animal non humain. La présente invention a également trait à un animal non humain portant l'échafaudage selon la présente invention.
PCT/IB2011/053586 2010-08-17 2011-08-11 Echafaudage polymère de polycaprolactone tridimensionnel Ceased WO2012023093A1 (fr)

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ITRM2010A000454 2010-08-17
ITRM2010A000454A IT1406053B1 (it) 2010-08-17 2010-08-17 Scaffold tridimensionale polimerico di policaprolattone

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Cited By (1)

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Publication number Priority date Publication date Assignee Title
WO2016054354A1 (fr) * 2014-10-02 2016-04-07 Dana-Farber Cancer Institute, Inc. Compositions et méthodes pour le traitement de tumeurs malignes

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WO2008041854A1 (fr) 2006-10-06 2008-04-10 Vrije Universiteit Medisch Centrum (Vumc) Enrichissement de cellules souches et/ou de cellules progénitrices
WO2009054006A2 (fr) 2007-10-26 2009-04-30 National Institute Of Immunology Structure de polymère biodégradable et procédé pour la préparer
WO2010067086A2 (fr) 2008-12-12 2010-06-17 The University Of Manchester Échafaudage de réparation tissulaire

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WO2016054354A1 (fr) * 2014-10-02 2016-04-07 Dana-Farber Cancer Institute, Inc. Compositions et méthodes pour le traitement de tumeurs malignes
US10365280B2 (en) 2014-10-02 2019-07-30 Dana-Farber Cancer Institute, Inc. Compositions and methods for treating malignancies
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