In vitro growth and proliferation of genetically modified multipotent neural stem cells and their progeny

A method of producing genetically modified neural cells comprising:

• (a) obtaining cells derived from juvenile or adult mammalian neural tissue containing at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons, astrocytes, and oligodendrocytes;
• (b) preparing a culture medium containing one or more predetermined growth factors capable of inducing multipotent neural stem cell proliferation;
• and
• (c) preparing a cell culture by combining the cells obtained in (a) with the culture medium prepared in (b) to induce proliferation of said multipotent neural stem cell to produce multipotent neural stem cell progeny which includes daughter multipotent neural stem cells and introducing exogenous DNA that does not contain oncogenes to genetically modify said multipotent neural stem cell progeny.

A method of producing genetically modified neural cells comprising:

• (a) obtaining cells derived from human neural tissue containing at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons, astrocytes, and oligodendrocytes;
• (b) preparing a culture medium containing one or more growth factors capable of inducing multipotent neural stem cell proliferation;
• and
• (c) preparing a cell culture by combining the cells obtained in (a) with the culture medium prepared in (b) to induce proliferation of said multipotent neural stem cell to produce multipotent neural stem cell progeny which includes daughter multipotent neural stem cells and introducing exogenous DNA that does not contain oncogenes to genetically modify said multipotent neural stem cell progeny.

A method of producing genetically modified differentiated neural cells comprising:

• (a) obtaining cells derived from mammalian neural tissue containing at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons, astrocytes, and oligodendrocytes, wherein said obtained cells have not been cultured in a serum-containing medium;
• (b) preparing a substantially serum-free culture medium containing one or more predetermined growth factors capable of inducing multipotent neural stem cell proliferation,
• (c) preparing a cell culture by combining the cells obtained in (a) with the culture medium prepared in (b) to induce proliferation of said multipotent neural stem cell to produce proliferating multipotent neural stem cells and multipotent neural stem cell progeny which includes daughter multipotent neural stem cells, and
• (d) subjecting the cells proliferated in (c) to culture conditions that induce neural cell differentiation to produce differentiating and differentiated neural cells, and introducing exogenous DNA that does not contain oncogenes to genetically modify said differentiated and/or differentiating neural cells to produce genetically modified differentiated neural stem cell progeny.