Method of producing native components, such as growth factors or extracellular matrix proteins, through cell culturing of tissue samples for tissue repair

9,220,803

12/443282

October 02, 2007

A medical composition is disclosed, which is injectable and which comprises a mixture of native components, which are obtainable by culturing one or more cell samples from a human or animal during normal conditions, said native components being included in the group consisting of growth factors, extracellular matrix proteins, and other substances produced by said cell samples during normal conditions, and a pharmaceutically acceptable carrier, as well as a method for producing the native components, a method for producing the medical composition, a method for treating a subject in need of tissue repair by injection of the medical composition, and use of said mixture of native components for the production of said medical composition for tissue repair via injection. A medical composition is disclosed, which is injectable and which comprises a mixture of native components, which are obtainable by culturing one or more cell samples from a human or animal during normal conditions, said native components being included in the group consisting of growth factors, extracellular matrix proteins, and other substances produced by said cell samples during normal conditions, and a pharmaceutically acceptable carrier, as well as a method for producing the native components, a method for producing the medical composition, a method for treating a subject in need of tissue repair by injection of the medical composition, and use of said mixture of native components for the production of said medical composition for tissue repair via injection.

Lidgren, Lars (Lund, SE)

1. A method for producing a mixture of native components, included in the group consisting of growth factors, extracellular matrix proteins and other substances produced by cell samples during normal conditions, consisting of the steps of: adding one or several different cell samples of human or animal origin to a bioreactor containing a nutrient medium, wherein a culturing medium is obtained; cell culturing during normal conditions, wherein the cell culturing is carried out under rotating condition; drawing off at least once the culturing medium including native components produced during the cell culturing; separating the native components from the culturing medium thereby obtaining a mixture of native components; and optionally, sterilizing, enriching, adding a carrier, and/or freeze-drying the separated native components; wherein the drawing off takes place continuously and the drawn-off culturing medium is replaced with new nutrient medium, said bioreactor having two or more interconnected chambers separated by a Millipore filter which allows for the culturing medium containing native components produced to diffuse into a second chamber or third chamber and so on and for separation of the cells from the culturing medium containing native components in the bioreactor, and wherein each cell sample originates from a tissue biopsy taken from cartilage and/or bone.

2. The method according to claim 1 , wherein the method further comprises freeze-drying the separated native components.

3. The method according to claim 1 , wherein a matrix is present during the cell culturing in order to increase the production of the native components.

4. The method according to claim 3 , wherein the matrix is chosen from starch beads, polymer beads, and beads of alginate, collagen, hyaluronic acid, and chitosan.

5. The method according to claim 1 wherein the mixture of native components is sterilized.

6. The method according to claim 1 , wherein the cell culturing is carried out during at least 7 weeks.

7. The method according to claim 1 , wherein the native components produced are separated from the culturing medium by size or weight or by affinity.

8. The method according to claim 7 , wherein the native components produced are separated from the culturing medium by centrifugation or are separated in a column.

9. The method according to claim 1 , wherein each of the separated native components is further enriched before freeze-drying.

10. The method according to claim 1 , wherein the cell sample is a cell line.

11. The method according to claim 10 , wherein the cell One is commercial.

12. The method according to claim 1 , wherein a carrier is added to the mixture of native components produced before or after an optional freeze-drying step.

13. The method according to claim 12 , wherein the carrier is chosen from the group consisting of natural or synthetic polymers, and ceramic materials.

14. The method according to claim 13 , wherein the carrier is chosen from hyaluronic acid, fibrin glue, chitosan, dextran, collagen, alginate, a biopolymer of different materials having different modules of elasticity, and a biopolymer with an elasticity gradient within the same material.

15. The method according to claim 1 , wherein the proportion between the native components and the carrier is about 1:10 parts by weight.

16. The method according to claim 1 , wherein said native components are growth factors and extracellular matrix proteins (GECM).

17. The method according to claim 16 , wherein the growth factors are selected from transforming growth factor, bone morphogenic protein (BMP-2), PTHrP, osteoprotegrin (OPG), basic fibroblast growth factor (bFGF), insulin-like growth factor, Indian hedgehog activator, an NE-kappa B ligand (RANKL), a vascular endothelial growth factor, and autologous growth factors; wherein the extracellular matrix proteins (ECM) are fibronectin, vitronectin, chondroadherin or aggrecans.

18. The method according to claim 1 , wherein the cell culturing is carried out in the absence of a three-dimensional matrix.

19. The method according to claim 1 , wherein said mixture of native components comprises a proteineous component parathyroid hormone, a prostaglandin, an osteoprotegrin, a sex steroid, or a cytokine.

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