Brain-derived neurotrophic factor, ciliary neurotrophic factor and their combination for in vitro differentiation of human umbilical blood-derived mesenchymal stem cells into nerve-like cells / 中国组织工程研究

ABSTRACT

BACKGROUND: Umbilical cord blood mesenchymal stem cells (UCBMSCs) have been shown to differentiate into nerve-like cells under the condition of in vitro culture. Brain-derived neurotrophic factors (BDNF) combined with ciliary neurotrophic factors (CNTF) at certain concentration can in vitro induce the differentiation of UCBMSCs into high proportion of neuronal-like cells. OBJECTIVE: To investigate the feasibility of BDNF, CNTF and their combination for in vitrodifferentiation of UCBMSCs into nerve-like cells.METHODS: UCBMSCs of passage 5 were induced to differentiate into nerve-like cells using 5,10, 20 ug/L BDNF, 5,10, 20 ug/L CNTF or their combination. A blank control group was set, with no any interventions. Cell morphology was observed under inverted phase contrast microscope. At 1, 3, and 6 days of experiment, immunocytochemical staining for neuron specific enolase and glial fibrillary acidic protein was performed. The proportion of differentiated neuronal-like cells and glial cell-like cells was calculated.RESULTS ANDCONCLUSION:After induced differentiation into nerve-like cells, human UCBMSCs exhibited changeable morphology with contracted cell body, enhanced refraction of nucleus, and dendrite- and axon-like structure. Compared with blank control group, BDNF and CNTF couldsignificantly enhance the differentiation proportion of UCBMSCs into nerve-like cells. 20 ug/LBDNF combined with 20 ug/L CNTF yielded highest differentiation proportion of human UCBMSCs into nerve-like cells. These findings indicate that human UCBMSCs can differentiate into nerve-like cells after in vitro induction of BDNF, CNTF or their combination.