Methods and compositions to facilitate repair of avascular tissue

1. A method for treating an avascular zone in a patient in need thereof, the method comprising:

an isolation and expansion process consisting of:

obtaining whole marrow from the patient in need thereof;

centrifuging the whole marrow to separate nucleated cells from red blood cells;

removing the nucleated cells from the centrifuged whole marrow;

washing and counting the nucleated cells;

culturing the nucleated cells in a culture medium under a selective pressure of about 1% to about 10% oxygen for 1-28 days, wherein the culture medium is maintained at a pH from 6.6 to 6.9 and the culture medium comprises 10% to 20% human platelet lysate;

selecting viable mesenchymal stem cells capable of growth in the culture medium under the selective pressure of about 1% to about 10% oxygen;

an implantation process comprising:

providing the selected, viable, mesenchymal stem cells for implantation in the avascular zone;

implanting platelet cell lysate or platelets prior to, during, or after implanting the selected, viable, mesenchymal stem cells;

wherein the implanting of the platelet cell lysate or platelets and the selected, viable, mesenchymal stem cells is in the avascular zone.

2. The method of claim 1 , wherein the culture medium has an osmolarity of about 350 mOsm to about 600 mOsm.

3. The method of claim 1 , wherein about 10 5 -10 9 selected, viable mesenchymal stem cells are implanted.

4. The method of claim 1 , wherein the implanting also includes, either separately or in combination, epidural supplements, growth factors, cytokines, integrins, and cadherins to facilitate blood flow.

5. The method of claim 1 , wherein the selected, viable mesenchymal stem cells are implanted into a degenerative intervertebral disc.

6. The method of claim 5 , wherein the selected, viable mesenchymal stem cells are implanted into a posterior disc annulus of the degenerative intervertebral disc.

7. A method for treating an avascular zone in a patient in need thereof, the method comprising:

an isolation and expansion process consisting of:

obtaining whole marrow from the patient in need thereof;

centrifuging the whole marrow to separate nucleated cells from red blood cells;

removing the nucleated cells from the centrifuged whole marrow;

washing and counting the nucleated cells;

culturing the nucleated cells in a culture medium under a selective pressure of about 10% oxygen and then culturing the nucleated cells under decreasing levels of oxygen until about 1% oxygen, wherein the culturing from about 10% oxygen to about 1% oxygen is for 1-28 days wherein the culture medium is maintained at a pH from 6.6 to 6.9 and the culture medium comprises 10% to 20% human platelet lysate;

selecting viable mesenchymal stem cells capable of growth in the culture medium under the selective pressure of about 10% to about 1% oxygen;

an implantation process comprising:

providing the selected, viable mesenchymal stem cells for implantation in the avascular zone;

implanting platelet cell lysate or platelets prior to, during, or after implanting the selected, viable mesenchymal stem cells;

wherein the implanting of the platelet cell lysate or platelets and the selected, viable mesenchymal stem cells is in the avascular zone.

8. The method of claim 7 , wherein the culture medium has an osmolarity of about 350 mOsm to about 600 mOsm.

9. The method of claim 7 , wherein about 10 5 -10 9 selected, viable mesenchymal stem cells are implanted.

10. The method of claim 7 , wherein the implanting also includes, either separately or in combination, epidural supplements, growth factors, cytokines, integrins, and cadherins to facilitate blood flow.

11. The method of claim 7 , wherein the selected, viable mesenchymal stem cells are implanted into a degenerative intervertebral disc.

12. The method of claim 11 , wherein the selected, viable mesenchymal stem cells are implanted into a posterior disc annulus of the degenerative intervertebral disc.

13. A method for treating an avascular zone in a patient in need thereof, the method comprising:

an isolation and expansion process consisting of:

obtaining whole marrow from the patient in need thereof;

centrifuging the whole marrow to separate nucleated cells from red blood cells;

removing the nucleated cells from the centrifuged whole marrow;

washing and counting the nucleated cells;

culturing the nucleated cells in a culture medium under a selective pressure of about 10% oxygen and then culturing the nucleated cells under decreasing levels of oxygen until about 5% oxygen, wherein the culturing from about 10% oxygen to about 5% oxygen is for 1-28 days, wherein the culture medium is maintained at a pH from 6.6 to 6.9 and the culture medium comprises 10% to 20% human platelet lysate;

selecting viable mesenchymal stem cells capable of growth in the culture medium under the selective pressure of about 10% to about 5% oxygen;

an implantation process comprising:

providing the selected, viable mesenchymal stem cells for implantation in the avascular zone;

implanting platelet cell lysate or platelets prior to, during, or after implanting the selected, viable mesenchymal stem cells;

wherein the implanting of the platelet cell lysate or platelets and the selected, viable mesenchymal stem cells is in the avascular zone.