Trophic stimulation of embryonic cells from rat corpus striatum

AM CRUZ GAITÁN; M REYNALDO; J TAU; NG CARRI

Huerta Grande, Cordoba

Congreso; IRCN; 2009

Reunion Conjunta del TN y SAN

Neural Stem Cells (NSC) from the sub ventricular zone of mammalian brain have become essential for neural regeneration research. Progenitors in the Corpus Striatum (CS) were discovered a few years ago, but little is known about their proliferation and differentiation. The purpose of this study was to analyze CS morpho-immune profiles using bioassays under basal and different trophic stimulation conditions. Briefly, primary cells obtained from CS of rat embryos at E13-14 were cultivated floating in DMEM-F12, bFGF, and B27.  After forming neurospheres they were then placed inside gelified collagen I, and cultured under basal conditions or with the addition of NGF, NT-3, or NTN. We obtained optimum growth of neurites that were measured in number and length after 24 and 48 hours. The expression of proliferation markers (PCNA, Ki67) and neural progenitor markers (GFAP, Nestin, Vimentin, O4, A2B5, Pax6, S100, TubIII, and NeuN) were also analyzed.  The initial behaviour of CS primary cell cultures showed distinguishable neurospheres, that when placed in collagen gels generated neurites, depending on the trophic support. Also, glial cells grew out from the neurospheres. Antibody staining was strongly positive with the exception of NeuN and O4 that were faint. The bioassay system allowed us to conclude that CS cells can generate neural progenitors useful for research on proliferation and differentiation. Our assay showed high reproducibility, short culture time and high resolution to trace neurites growing out from neurons in a few hours or visualize glial cells outgrowth.