Induced Differentiation Of Human Umbilical Cord Blood Monocytes Into Neuron-Like Cells In Vitro

Objective: To explore the isolation, culture , purification and expansion of human umbilical cord blood mesenchymal stem cells (MSCs) into neuron-like cells in vitro. And the effect of bFGF on cells differentiation. To probe the condition how mesenchymal stem cells in human umbilical cord blood can be differentiated into neuron-like cells in vitro, so as to offer the technology and theory for treatment of spinal cord injury in the future.Methods: Human mononuclear cells (MNCs) obtained from umbilical cord blood with density gradient centrifugation were planted into culture fluid named H-DMEM which contains fetal bovine serum 20%. Then the MNCs were static cultured in incubator under the condition of 37℃, 5%CO2 and saturated humidity , and bFGF was added into some culture flasks which are called aim group after 72h, final concentration of bFGF is 10ng/ml. Observe cells growth and the effect of bFGF with inverted microscope. Choice the mononuclear cells which were cultured in vitro after 3 days, 7 days and 13 days. Then detect the cell surface marker of nestin, MAP2 and GFAP by immunohistochemistry method. Observe positive cells in 10 nonoverlapping fields of vision by light microscope. Then calculate percentage of positive cells of nestin, MAP2 and GFAP, and the positive percentage is recorded as ( x ±s) %.Nestin, MAP2 and GFAP monoclonal antibodies were replaced by 0.01mol/L PBS in negative control group. Use PBS that contains 20g/L bovine serum albumin wash the first filial generation MSCs, then...