Claims
- 1. A method of regenerating skeletal tissue of a subject comprising the steps of:
- a culturing a proliferating cell population comprising autologous bone marrow stroma cells;
- b. manipulating them in suspension above soft agar at a concentration in excess of 2.times.10.sup.6 cell/ml of medium for at least five days;
- c. harvesting the cells;
- d. embedding the cells at a concentration in excess of 80.times.10.sup.6 cells/ml of a biological resorbable immobilization vehicle (BRIV) including in excess of 10% serum, 100 mg/ml fibrinogen, and 60 units/ml thrombin in an excess of 40mM CaC1.sub.2 and 2,000 units of aprotonin;
- e. removing a damaged part of the subject's skeletal tissue to leave a clean geometric shaped defect; and
- f. filling the defect with an implant at least including the combined cell concentration and BRIV.
- 2. The method as set forth in claim 11 above, wherein the BRIV includes 15-30% serum and 60 units/ml thrombin in an excess of 60mM CaC1.sub.2.
- 3. The method as set forth in claim 1 above, wherein the biological implant is stored under low temperature conditions after preparation with a preservative medium containing 90% F.C.S. and 10% DMSO, and wherein there is about 20% serum.
- 4. The method as set forth in claim 1 above, wherein the harvested cells are stored under low temperature conditions after preparation with a preservative medium containing 90% F.C.S. and 10% DMSO, and embedded at a later date in BRIV.
- 5. The method as set forth in claim 1 above, wherein the skeletal tissue to be repaired comprises articular cartilage and the joint operated on is subjected to a continuous passive motion after implantation.
- 6. The method according to claim 1, wherein the serum is selected from the group consisting of fetal calf serum, umbilical cord serum from the second trimester, and horse serum.
- 7. A method of regenerating skeletal tissue of a subject, comprising the steps of:
- a. culturing a proliferating cell population comprising autologous muscle fibroblast derived chrondocytes;
- b. manipulating the cells in suspension and culturing the cells above soft agar at a concentration in excess of 2.times.10.sup.6 cells/ml of medium for at least 3 days;
- c. harvesting the cells;
- d. embedding the cells at a concentration in excess of 80.times.10.sup.6 cells/ml of a biological resorbable immobilization vehicle (BRIV), the BRIV comprising in excess of 10% serum, 100 mg/ml fibrinogen, 60 units/ml thrombin in an excess of 40mM CaC1.sub.2, and 2000 units of aprotonin;
- e. removing a damaged part of the subject's skeletal tissue to leave a clean geometric shaped defect; and
- f. filling the defect with an implant at least including the combined cell concentration and BRIV.
- 8. The method of claim 7 as set forth above, wherein the BRIV includes 15-30% serum and 60 units/ml thrombin in an excess of 60mM CaC1.sub.2.
- 9. The method as set forth in claim 7, wherein the biological implant is stored under low temperature conditions after preparation with a preservative medium containing 90% FCS and 10% DMSO, and wherein there is about 20% serum.
- 10. The method as set forth in claim 7 above, wherein the harvested cells are stored under low temperature conditions after preparation with a preservative medium containing 90% FCS and 10% DMSO, and embedded at a later date in BRIV.
- 11. The method as set forth in claim 7 above, wherein the skeletal tissue to be repaired comprises articular cartilage and bone.
- 12. A method of regenerating skeletal tissue in a person, comprising the steps of:
- a. obtaining a bone marrow sample from a donor;
- b. culturing a proliferative cell population comprising bone marrow osteogenic-chondrogenic progenitor cells;
- c. harvesting the cells;
- d. manipulating the cells in suspension above soft agar at a concentration in excess of 2.times.10.sup.6 cells/ml of medium for at least five days;
- e. embedding the cells at a concentration in excess of 80.times.10.sup.6 cells/ml of a biological resorbable immobilization vehicle (BRIV) including in excess of 10% serum, 100 mg/ml fibrinogen and 60 units/ml thrombin in 60 mM CaC1.sub.2 and 2000 units of aprotonin;
- f. removing a damaged part of the person's skeletal tissue to leave a clean geometric shaped defect; and
- g. filling the defect with an implant at least including the combined cell concentration and BRIV.
- 13. The method of claim 12 wherein the bone marrow sample is obtained by aspiration from the iliac crest of the donor.
- 14. The method of claim 12 wherein the donor of the bone marrow sample and the person receiving the implant are the same person.
- 15. The method as set forth in claim 12 above wherein the BRIV includes 15-30% serum and 60 units/ml thrombin in an excess of 60 mM CaC1.sub.2 and 2000 units of aprotonin.
- 16. The method as set forth in claim 12 above, wherein the biological implant is stored under low temperature conditions after preparation with a preservative medium containing 90% F.C.S. and 10% DMSO.
- 17. The method as set forth in claim 12 above, wherein the harvested cells are stored under low temperature conditions after preparation with a preservative medium containing 90% F.C.S. and 10% DMSO, and embedded at a later date in BRIV.
- 18. The method as set forth in claim 12 above, wherein the skeletal tissue to be repaired comprises articular cartilage and the joint operated on is subjected to a continuous passive motion after implantation.
- 19. The method according to claim 12, wherein the serum is selected from the group consisting of fetal calf serum, umbilical cord serum from the second trimester, and horse serum.
- 20. The method as set forth in claim 12 wherein the cells and the BRIV are embedded in a suitably shaped biodegradable bone substitute.
- 21. The method as set forth in claim 12 above wherein the concentration of the cells is between 80.times.10.sup.6 cells/ml and 160.times.10.sup.6 cells/ml BRIV.
- 22. The composition of claim 12, wherein the protease inhibitor is selected from the group consisting of polysaccharides, plasma protease, synthetic and natural non-plasma protease inhibitors.
- 23. The method as set forth in claim 12 wherein the cells originate from a nonhuman source.
CROSS-REFERENCE
This is a continuation-in-part application of copending U.S. Serial No. 187,730, filed on April 29, 1988, pending, which is incorporated by reference herein.
US Referenced Citations (1)
Number |
Name |
Date |
Kind |
4846835 |
Grande |
Jul 1989 |
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Continuation in Parts (1)
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Number |
Date |
Country |
Parent |
187730 |
Apr 1988 |
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