Autophagy-related Gene Beclin-1Expression In Neuron-like Differentiation Of Human Bone Marrow Mesenchymal Stem Cells

BACKGROUND: Bone marrow mesenchymal stem cells have potential toself-renewal and multi-lineage differentiation. But after a long period ofculture in vitro, the proliferation and differentiation capacities of bonemarrow mesenchymal stem cells gradually loss, the mechanism underlyingwhich is not clear now.OBJECTIVE:1. Isolate and culture human bone marrow mesenchymal stem cells in vitro.2. Using basic fibroblast growth factor andepidermal growth factor topromote the differentiation from human bone marrow mesenchymal stemcells into neuron-like cells in vitro.3. To observe the expression of autophagy-related gene Beclin-1indifferentiation from human bone marrow mesenchymal stem cells intoneuron-like cells in vitro.METHODS:1.Using Ficoll-Paque to Isolate human mononuclear cells under sterileconditions.And culture human bone marrow mesenchymal stem cells in vitro.2.The changes of morphological characteristics of neuron-like cellsdifferentiated from human bone marrow mesenchymal stem cells induced byepidermal growth factor were observed. 3.The expression of neuron-specific enolase in treated and untreated humanbone marrow mesenchymal stem cells were detected usingimmunocytochemistry.4.The Beclin-1protein expressions were detected by western blot before andafter induction.RESULTS AND CONCLUSION:1.Human bone marrow mesenchymal stem cells can survive a certain time invitro, and can accept a certain degree of pretreatment in vitro environment.2.After being induced, human bone marrow mesenchymal stem cellspresented classical neuron-like morphology; the expressions ofneuron-specific enolase were78.7%, respectively.3.The expression of Beclin-1protein was changed correspondingly during theinduction, which increased after30minutes of induction and decreasedgradually after1hour of induction. Human bone marrow mesenchymal stemcells could be induced into neuron-like cells in vitro by epidermal growthfactor. Autophagy-related gene was highly expressed in the induction of earlydifferentiation and the expression gradually reduced until it remained at a lowlevel during the differentiation.