Method for in vitro production of three-dimensional vital cartilage tissue and use thereof as transplant material
AssigneeCo.don Aktiengesllschaft
InventorsJeanette Libera, Ursula Anderer, Karl-Gerd Fritsch, Olivera Josimovic-Alasevic
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Abstract
The invention relates to a remarkably simple method for the in vitro production of three-dimensional, vital and mechanically stable cartilage or bone tissue and to the use thereof as a transplantation material for treating cartilage or bone defects and degenerative diseases such as rheumatism or arthrosis, and to the use thereof in testing active substances and physical factors. The invention is also directed to the cartilage or bone tissue and therapeutical formulations produced thereby, e.g. injection solutions comprising such tissue.
Claims17
A method for treating a defect in cartilage tissue of a patient, the method consisting of:
- (a) isolating cartilage cells from a human or animal organism,
- (b) subjecting said cells to stationary suspension culture in a cell culture vessel having a hydrophobic surface and tapered bottom to form a cell aggregate in the form of a spheroid,
- wherein said culturing is performed in a cell culture medium consisting of:
- (i) DMEM, or DMEM and HAMS, and
- (ii) a serum chosen as one of autogenous, xenogeneic and allogenic with respect to the patient, and
- wherein said spheroid comprises at least 40% by volume of extracellular matrix (ECM), differentiated cells embedded within the ECM, has an outer region comprising cells capable of proliferation and migration, and has a size defined as one of:
- (i) a diameter of 500-700 μm,
- (ii) containing 1×104 to 1×107 cells, or
- (iii) a size that may be produced in a well of a 96-well cell culture plate,
- (c) repeating steps (a) and (b) to obtain a plurality of spheroids as a transplant material, or, alternatively,
- (c′) repeating steps (a) and (b) to form at least two spheroids, and then fusing the at least two spheroids by culturing the at least two spheroids together according to (b) to obtain a single spheroid as a transplant material, and
- (d) transplanting the transplant material into the defect of the patient,
- wherein between (a) and (b) the cells may optionally be grown in a monolayer culture, or in a case in which the cell culture vessel has a non-hydrophobic surface, between (a) and (b) the surface of the cell culture vessel can be rendered hydrophobic by coating the surface with agar or agarose.
A method for making cartilage tissue transplant material to treat a defect in cartilage tissue of a patient, the method consisting of:
- (a) subjecting cartilage cells isolated from a human or animal organism to stationary suspension culture in a cell culture vessel having a hydrophobic surface and tapered bottom to form a cell aggregate in the form of a spheroid,
- wherein said culturing is performed in a cell culture medium consisting of:
- (i) DMEM, or DMEM and HAMS, and
- (ii) a serum chosen as one of autogenous, xenogeneic and allogenic with respect to the patient, and
- wherein said spheroid comprises at least 40% by volume of extracellular matrix (ECM), differentiated cells embedded within the ECM, has an outer region comprising cells capable of proliferation and migration, and has a size defined as one of:
- (i) a diameter of 500-700 μm,
- (ii) containing 1×104 to 1×107 cells, or
- (iii) a size that may be produced in a well of a 96-well cell culture plate, and
- (b) repeating step (a) to obtain a plurality of spheroids as the cartilage tissue transplant material, or, alternatively,
- (b′) repeating step (a) to form at least two spheroids, and then fusing the at least two spheroids by culturing the at least two spheroids together according to step (a) to obtain a single spheroid as the cartilage tissue transplant material,
- wherein the cartilage tissue transplant material is produced in vitro and is three-dimensional vital cartilage tissue of a different size or shape than a defect of a patient.
A method of treating a cartilage defect in a patient, comprising transplanting into said defect a transplant material comprising a plurality of cartilaginous spheroids,
- wherein each of said spheroids comprises at least 40% by volume of extracellular matrix (ECM), differentiated cells embedded within the ECM, has an outer region comprising cells capable of proliferation and migration, and has a size defined as one of:
- (i) a diameter of 500-700 μm,
- (ii) containing 1×104 to 1×107 cells, or
- (iii) a size that may be produced in a well of a 96-well cell culture plate,
- wherein the transplant material is one of autogenous, xenogeneic or allogeneic with respect to the patient, and
- wherein the transplant material comprises a number of spheroids which is insufficient to initially fill the defect, but which will grow to sufficiently fill the defect over time, irrespective of the shape or size of the defect.
Dependencies are inferred from the claim text.