Osteoblasts derived from oral neuroectodermal stem cells and their use in jaw repair
The present invention concerns a method for inducing differentiation of neuroectodermal oral stem cells, in particular from FCS osteogenic medium PL osteogenic medium gingival tissue (GSCs), into osteoblasts by culturing them in an optimal serum-free medium supplemented by necessary components such as platelet lysate, growth hormone, heparin, and/or growth factors. The invention method provides osteoblasts for cell therapy, particularly for the restoration of bone defects in maxillary bones.
1 . A method for inducing differentiation of neuroectodemic oral stem cells into osteoblasts, said method comprising:
(a) a step of cell proliferation of neuroectodermal oral stem cells in a serum-free base culture medium supplemented with platelet lysate (PL) and human growth hormone (GH);
b) an osteoblastic differentiation step of the cell culture resulting from step a) in a serum-free base culture medium supplemented with a platelet lysate (PL) and at least one osteoblastic differentiation factor.
2 . Method according to claim 1 , wherein the culture of step a) is carried out up to a confluence of 80%.
3 . Method according to claim 1 , wherein the culture of step b) is carried out during 21 to 28 days.
4 . Method according to claim 1 , wherein the culture medium of steps a) and b) is further supplemented with heparin.
5 . Method according to claim 4 , wherein the culture medium of step a) contains from 1-2IU/ml of heparin, and/or the culture medium of step b) contains from 0.1-0.6 IU/ml of heparin.
6 . Method according to claim 1 , wherein said at least one osteoblastic differentiation factor is chosen from the group consisting of corticosteroids and glycerol phosphoric esters.
7 . Method according to claim 6 , wherein the said at least one differentiation factor is dexamethasone and/or b-glycerophosphate.
8 . Method according to claim 1 , wherein the neuroectoderm is oral stem cells are derived from gingival tissue.
9 . Cell population comprising osteoblasts produced by the method according to claim 1 , said osteoblasts expressing the homeogen Msx2.
10 . Cell population as defined in claim 9 , for use in cell therapy.
11 . Cell population for use according to claim 10 , for the restoration of bone defects of maxillary bones.
12 . Culture medium for the proliferation of oral neuroectodermal stem cells comprising a serum-free base culture medium supplemented with platelet lysate (PL) and human growth hormone (GH), and optionally heparin at a concentration ranging from 1-2 IU/ml.
13 . Culture medium for the differentiation of neuroectodermal oral stem cells into osteoblasts comprising a serum-free base culture medium supplemented with a platelet lysate (PL) and at least one osteoblastic differentiation factor, and optionally heparin at a concentration ranging from 0.1 -0.6 IU/ml.
14 . Culture medium according to claim 13 , wherein said at least one osteoblastic differentiation factor is chosen from the group consisting of corticosteroides and glycerol phosphoric esters.
15 . Culture medium according to claim 14 , wherein said at least one osteoblastic differentiation factor is dexamethasone and/or b-glycerophosphate.