Schwann cells originating in myeloid interstitial cells
There is provided a method of inducing bone marrow stromal cells to differentiate into bone marrow stromal cell-derived Schwann cells in vitro, comprising the steps of: collecting bone marrow stromal cells from bone marrow and culturing the cells in a standard essential culture medium supplemented with a serum; adding a reducing agent to the culture medium and further culturing the cells; adding a differentiation inducing agent to the culture medium and further culturing the cells; and adding a cyclic AMP-augmenting agent or a cyclic AMP analogue and/or a glial cell differentiation and survival stimulating factor to the culture medium, and further culturing the cells to obtain the bone marrow stromal cell-derived Schwann cells. There are also provided bone marrow stromal cell-derived Schwann cells obtained thereby and a pharmaceutical composition for neural regeneration that comprises them.
1. A method of inducing bone marrow stromal cells to differentiate into bone marrow stromal cell-derived Schwann cells in vitro, comprising: (1) collecting bone marrow stromal cells from bone marrow, and culturing said cells in a standard essential culture medium supplemented with a serum (2) adding a reducing agent to said culture medium, and further culturing said cells; (3) adding retinoic acid to said culture medium, and further culturing said cells; and (4) adding forskolin, and/or a differentiation, survival and growth stimulating factor which acts on nerves and glial cells to said culture medium, and further culturing said cells to obtain said bone marrow stromal cell-derived Schwann cells.
2. The method as defined in claim 1 , wherein said standard essential culture medium is an Eagle's alpha modified minimum essential medium.
3. The method as defined in claim 1 , wherein said serum is fetal calf serum.
4. The method as defined in claim 1 , wherein said reducing agent is sulfhydryl-containing reagents.
5. The method as defined in claim 4 , wherein said sulfhydryl-containing is β-mercaptoethanol.
6. The method as defined in claim 1 , wherein the concentration of said reducing agent is between 1 nM and 10 mM.
7. The method as defined in claim 1 , wherein the culturing time in step (2) is between 1 hour and 5 days.
8. The method as defined in claim 1 , wherein the concentration of said retinoic acid is between 0.001 ng/ml and 1 μg/ml.
9. The method as defined in claim 1 , wherein the culturing time in step (3) is within 30 days.
10. The method as defined in claim 1 , wherein the concentration of said forskolin is between 0.001 ng/ml and 100 μg/ml.
11. The method as defined in claim 1 , wherein said glial cell differentiation and survival stimulating factor is selected from the group consisting of neuregulin, platelet-derived growth factor-AA, basic fibroblast growth factor, and mixtures thereof.
12. The method as defined in claim 11 , wherein said neuregulin is Heregulin, a subtype of the same.
13. The method as defined in claim 1 , wherein the concentration of said glial cell differentiation and survival stimulating factor is between 0.001 ng/ml and 100 μg/ml.
14. The method as defined in claim 1 , wherein the culturing time in step (4) is within 30 days.