Differentiation of mesenchymal cells derived from mouse fetal liver into neuronal cells in vitro / 第二军医大学学报

ABSTRACT

Objective:To investigate potency of mouse fetal liver mesenchymal cells differentiating into neural cells in vitro .Methods: Sterile cells from 14.5 day old mouse fetal liver were cultured in DMEM/HEPES/F12 supplemented with 20%FCS and adherent cells were acquired after discarding nonadherent cells. The 5th passage cells were induced by ? mercaptoethanol (? ME) and all trans retinoic acid (ATRA). The characteristics of treated cells were assayed by immunocytochemistry staining and semi quantitative RT PCR method on 5 h and 5 d after induction. Results: After induction by ? ME and ATRA, approximately 80% of the cells exhibited typical neuronal morphology;most cells expressed neuronal phenotype such as neuron specific nuclear protein(NeuN), neuron specific tubulin Ⅲ(TuJ 1), neurofilament M(NF M),and neuron specific enolase(NSE).However, microtubule associated protein tau(Tau 1)and microtubule associated protein 2 (MAP 2) as indicators of mature neuronal phenotype were undetectable. Semi quantitative RT PCR showed increased mRNA expression of brain factor 1(BF 1), brain 3(Brn 3), and neurofilament L(NF L) in treated cells versus untreated cells ( P