IMBICE   05372
INSTITUTO MULTIDISCIPLINARIO DE BIOLOGIA CELULAR
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
ACTIVATION OF NEURAL STEM CELLS TO STRONG DIFFERENTIATION AND VIGOROUS OUTGROWTH BY TROPHIC FACTORS TANDEM INSIDE COLLAGEN GEL
Autor/es:
CRUZ GAITÁN ANA MARIA; REYNALDO M; CARRI NÉSTOR GABRIEL
Lugar:
Cancun
Reunión:
Workshop; IBRO Whorkshop for Latinamericans Fellows; 2012
Institución organizadora:
IBRO
Resumen:
Neural stem cells (NSC) from the sub ventricular zone brains in mammals have become quite significant for neural regeneration research. Progenitors in the Corpus Striatum (CS) were discovered but it is still under analysisi in vitro their proliferation and differentiation. Thus, the purpose of this study was to analyze CS cells in our model. under basal conditions (DMEM-F12, bFGF, hEGF , B27, 37°C, 100% humidity, and 5% CO2) and subsequent trophic stimulation (NGF, NT-3, NTN, FGF9 or Noggin) for  genic tandem activation of progenitors cells to induce differentiation and outgrowth. The model is based in 3-D collagen gelated bioassay after allowing the primary cells (obtained from CS of E14 rat embryos) their floatation in culture for 48h before an aliquot of the clumped cells is placed into gel to submit to trophic factors action.  After 72h we established a second passage using  dissociated neurospheres and cultured them during 48h lo allow new clump formation. We repit the same procedure described before.with trophic factors inside collagen gel during 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) was analyzed by immunocytochemistry techniques. As results CS culture showed clumped cells and clear neurospheres formation which still is a main characteristic to identify NSC. In our  3-D-bioassay, the majority of trophic stimulated cells generated a large number of long neurites emitted, founding clear differences respect to no treated ones. The expression of antigens was strongly positive for antibodies with the exception of NeuN and O4 that showed unclear soft mark. Our 3-D bioassay allowed us to conclude that CS cells have a capacity to permit differentiation and outgrowth that could be identified by markers and the inmunoprofile as neurons and glial cells.