Preparation of pure cultures of post-mitotic human neurons
AssigneeThe Trustees of the University of Pennsylvania
InventorsVirginia Lee, Samuel Pleasure
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Abstract
NTera 2/cl.D1 (NT2) cells, a human teratocarcinoma cell line, were manipulated following retinoic acid (RA) treatment to yield>95% pure cultures of neuronal cells (NT2-N cells). This culture method is capable of yielding sufficient highly differentiated post-mitotic NT2-N cells for both biochemical and molecular biological studies. NT2 cells can be transfected efficiently and the transfected gene products can be expressed in both NT2 and NT2-N cells.
Claims3
A method for producing a stable at least 95% pure population of post-mitotic human neurons expressing exogenous gene products comprising:
- transfecting at least one plasmid including a selectable marker into cultured NTera2/D1 cells;
- culturing said NTera2/D1 cells with retinoic acid to obtain a multi-layer culture;
- dispersing said cultured cells;
- and
- culturing said dispersed cells with cytosine arabinoside, fluorodeoxyuridine and uridine.
A method for preparing an at least 95% pure culture of post-mitotic human neurons comprising:
- culturing NTera2/D1 cells with retinoic acid to obtain a multi-layered culture;
- dispersing said cultured cells;
- culturing said dispersed cells with cytosine arabinoside, fluorodeoxyuridine and uridine.
Dependencies are inferred from the claim text.