Process of inducing differentiation of embryonic cell to cell expressing neural surface marker using OP9 or PA6 cells
View Patent ↗A method for inducing differentiation of an embryonic stem cell into an ectodermal cell and an ectoderm-derived cell, which comprises culturing the embryonic stem cell under non-aggregation conditions; a medium and a medium supernatant used in the method; an agent for inducing differentiation used in the method; a stroma cell or a stroma cell-derived factor having activity of inducing differentiation in the method; an antibody which specifically recognizes the stroma cell; an antigen which recognizes the antibody; a cell induced by the method; a method for evaluating or screening a substance relating to the regulation in a differentiation step from an embryonic stem cell into an ectodermal cell or an ectoderm-derived cell by carrying out the method; and a medicament comprising the stroma cell, the stroma cell-derived cell, the antibody, the antigen or the cell.
1. A method for producing a cell expressing a neural crest marker or a neural tube marker, comprising:
culturing under serum-free conditions a mammalian embryonic stem cell in vitro in the absence of retinoic acid and in the presence of a stroma cell without forming embryoid body, wherein the stroma cell is OP9 cell or PA6 cell.
2. A method for producing a dopaminergic neuron, an acetylcholinergic neuron, a γ-aminobutyratergic neuron or a serotonergic neuron, comprising:
culturing under serum-free conditions a mammalian embryonic stem cell in vitro in the absence of retinoic acid and in the presence of a stroma cell without forming embryoid body, wherein the stroma cell is OP9 cell or PA6 cell.
3. A method for producing a neural stem cell which is stained by an anti-nestin antibody comprising:
culturing under serum-free conditions a mammalian embryonic stem cell in vitro in the absence of retinoic acid and in the presence of a stroma cell without forming embryoid body, wherein the stroma cell is OP9 cell or PA6 cell.
4. The method according to any one of claim 1 , 2 or 3 , wherein the stroma cell is a stroma cell whose proliferation potency is deleted by a physicochemical treatment.
5. The method according to any one of claim 1 , 2 or 3 , wherein the stroma cell is a stroma cell whose proliferative potency is deleted by an antitumor agent, irradiation or pathologic tissue fixative.
6. The method according to claim 4 , wherein the physiocochemical treatment is an antitumor agent selected from the group consisting of mitomycin C, 5-fluorouracil, adriamycin and methotrexate.
7. The method according to any one of claim 1 , 2 or 3 , wherein the stroma cell is a stroma cell whose proliferative potency is deleted by a microwave fixation, a rapid freeze-substitution fixation, a glutaraldehyde fixation, a p-formaldehyde fixation, a formalin fixation, an acetone fixation, a Van fixation, a periodic acid fixation, a methanol fixation or an osmic acid fixation.
8. The method according to any one of claim 1 , 2 or 3 , wherein the stroma cell is an M-CSF deficient mouse calvaria-derived OP9 cell.
9. The method according to claim 8 , wherein the stroma cell is recognized by a monoclonal antibody produced by hybridoma FERM BP-7573.
10. The production method according to claim 1 , wherein the method produces a cell expressing said neural crest marker.
11. The production method according to claim 1 , wherein the method produces a cell expressing said neural tube marker.
12. The production method according to claim 2 , wherein the method produces said dopaminergic neuron.
13. The production method according to claim 2 , wherein the method produces said acetylcholinergic neuron.
14. The production method according to claim 2 , wherein the method produces said γ-aminobutyratergic neuron.
15. The production method according to claim 2 , wherein the method produces said serotonergic neuron.
16. The method according to any one of claims 1 , 2 or 3 , wherein the stroma cell is a mouse calvaria-derived MC3T3-G2/PA6 cell.
17. The method according to claim 8 , wherein the stroma cell is recognized by a monoclonal antibody produced by hybridoma FERM BP-7573.
18. The method according to claim 16 , wherein the stroma cell is recognized by a monoclonal antibody produced by hybridoma FERM BP-7573.