Method of inducing differentiation of bone marrow stromal cells to neural cells or skeletal muscle cells by introduction of Notch gene

Dopaminergic neurons produced by a method comprising the steps of:

• (1) isolating bone marrow stromal cells (BMSCs) from bone marrow, and culturing said cells in a standard essential culture medium supplemented with a serum;
• (2) introducing, into said cells, a nucleic acid comprising Notch sequences, wherein said Notch sequences consist of sequences encoding a Notch intracellular domain, and further culturing said cells;
• (3) adding a cyclic adenosine monophosphate (cAMP)-increasing agent or a cAMP analogue, and/or a cell differentiation stimulating factor to said culture medium, and further culturing said cells to produce a cell culture comprising neural cells;
• (4) culturing the resultant cell culture from Step (3) in a standard essential culture medium supplemented with a serum;
• and
• (5) adding glial derived neurotrophic factor (GDNF), and a cAMP-increasing agent or a cAMP analogue, and/or a cell differentiation stimulating factor other than said GDNF to said culture medium, and further culturing said cell culture to obtain a culture comprising said dopaminergic neurons.

Neural precursor cells produced by a method comprising the steps of:

• (1) isolating bone marrow stromal cells (BMSCs) from bone marrow, and culturing said cells in a standard essential culture medium supplemented with a serum;
• and
• (2) introducing, into said cells, a nucleic acid comprising sequences encoding a Notch intracellular domain, and further culturing said cells to obtain a culture comprising neural precursor cells;
• wherein the cells into which the nucleic acid has been introduced do not express a Notch extracellular domain.

Neural cells produced by a method comprising the steps of:

• (1) isolating bone marrow stromal cells (BMSCs) from bone marrow, and culturing said cells in a standard essential culture medium supplemented with a serum;
• (2) introducing, into said cells, a nucleic acid comprising sequences encoding a Notch intracellular domain, and further culturing said cells, wherein the cells into which the nucleic acid has been introduced do not express the Notch extracellular domain;
• and
• (3) adding a cyclic adenosine monophosphate (cAMP)-increasing agent or a cAMP analogue, and/or a cell differentiation stimulating factor to said culture medium, and further culturing said cells to produce a culture comprising said neural cells.

Dopaminergic neurons produced by a method comprising the steps of:

• (1) isolating bone marrow stromal cells (BMSCs) from bone marrow, and culturing said cells in a standard essential culture medium supplemented with a serum;
• (2) introducing, into said cells, a nucleic acid comprising sequences encoding a Notch intracellular domain, and further culturing said cells, wherein the cells into which the nucleic acid has been introduced do not express the Notch extracellular domain;
• (3) adding a cyclic adenosine monophosphate (cAMP)-increasing agent or a cAMP analogue, and/or a cell differentiation stimulating factor to said culture medium, and further culturing said cells to produce a cell culture comprising neural cells;
• (4) culturing the resultant cell culture from Step (3) in a standard essential culture medium supplemented with a serum;
• and
• (5) adding glial derived neurotrophic factor (GDNF), and a cAMP-increasing agent or a cAMP analogue, and/or a cell differentiation stimulating factor other than said GDNF to said culture medium, and further culturing said cell culture to obtain a culture comprising said dopaminergic neurons.