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Charakterizace míšních progenitorových buněk in vitro
Vitásková, Martina ; Klíma, Jiří ; Vodička, Petr ; Motlík, Jan
We performed in vitro studies with the spinal cord of mice and miniature pig. Employing papain dissociation system we isolated neural progenitor cells and cultured them in DMEM/F12 HEPES, Ala-Glu, gentamicin, heparin, B27 supplement without vitamin A, N2 supplement, EGF (Epidermal Growth Factor) and bFGF (Basic Fibroblast Growth Factor). We added retinoic acid and 10% fetal bovine serum into cultivation media to induce cell differentiation. Using immunocytochemistry we confirmed cell capability to differentiate into neurons (βIII-tubulin, Map 2A - Microtubule Associated Protein), astroglia (GFAP – Glial Fibrillary Acidic Protein), and oligodendroglia (CNPase - 2',3'-cyclic nucleotide 3'-phosphodiesterase). Cells can keep non-differentiated status for a longer period of time when cultured in low adhesion culturing flask or by addition of LIF (Leukemia Inhibitory Factor) to the cultivation media.
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