Differentiation Of Human Umbilical Cord Blood Mesenchymal Stem Cells Into Neural Cells Induced By Serum-free Conditioned Medium Of OECs

Objective:To study the possibility and mechanism by which the human umbilical cord blood mesenchymal stem cells (MSCs) are induced to differentiate into neural cells in vitro by serum-free conditioned medium of olfactory ensheasing cells (OECs) from neonatal rat, and to study their electrophysiological properties after neural differentiation.Methods:Primary culture and purification of neonatal rat OECs in vitro with Nash differential adhesion and cytarabine inhibition method, extract serum-free condition medium of OECs when it is at the best situation. Human umbilical cord blood was collected from full term deliveries scheduled for cesarean section, all specimens were obtained sterilely with preservative-free heparin, the mesenchymal stem cells were isolated by lymphocyte separation medium, purified by Nash differential adhesion and amplificated to passage 3,and marking as Human UCB-MSCs-P3. The study designed experimental group and control group. The UCB-MSCs-P3 were cultured with DMEM and the serum-free conditioned medium of OECs in the experimental group, and without the serum-free conditioned medium of OECs in the control group. At various time points (Oh,12h,24h,72h, lweek, 10days and 2weeks post-induced),BCB-MSCs were processed for morphology and immunocytochemistry, then patch clamp was used to detect the electrophysiological properties of diferentiated neuron-like cells at 14th-day post-induced.Results:The OECs could be purified to about 95 percent purity by the method. UCB-MSCs displayed neuronal morphologies and there were long processes out from the cell body when induced by serum-free conditioned medium of OECs in the experimental group, and maintain appeared Neuron-like cells after 2 weeks following induction. so as the number of immunocytochemistry cells labeled by neural markers NSE GFAP and nestin. Some UCB-MSCs-P3 that after induced displayed tetrodotoxin(TTX) sensitive and quickly activated Na+ current, and tetraethylammonium chloride(TEA) sensitive and slow activated K+ current as well as slow inward calcium current. However, the differentiated UCB-MSCs showing various morphological characteristics in the control group.Conclusions:①Serum-free conditioned medium of OECs can obviously induced human umbilical cord blood mesenchymal stem cells to differentiate into neural cells②The differentiated neurons have active electrophysiological properties as the mature neurons.