Study On Transdifferentiation Of Human Umbilical Cord Blood Derived Mesenchymal Stem Cells (HUCB-MSCs) Into Dopaminergic Neurons In Vitro

0bjectives To establish an induction system for HUCB-MSCs differentiating into dopaminergic neurons and exploit the potential therapeutic cell type for Parkinson's disease.Methods HUCB-MSCs of passage 5 were induced to differentiate into dopaminergic neural like cells using three different methods respectively:the antioxidate chemical method,the neurotrophic factor method and the chemical combined with neurotrphic factor combining method.Morphological changes were monitored during the differentiating procedure.The cells were then fixed with 4% paraformaldehyde and stained for neural specific marker Nestin,NSE,TH,DAT,DAD2 and glial marker GFAP.Real Time-PCR was performed to analyze the TH mRNA,DAT mRNA,and DRD2 mRNA expression in control and the induction group.Results HUCB-MSCs are characterized morphologically by a fibroblast-like population of cells before induction.The cells looked like neuronoid cells with taper body,many long tuber,strong three-dimensiona and cytoplast shrinking to the mucleus and became denser and more refracting after being incubated in the preinduction medium in 24 hours.After the differentiated induction,the cytoplasts kept shrinking to the nucleus and became round sharp,irregular taper and triangollar.Some cytoplasts have many furcal tubers similar to neuron. Immunohistochemistry showed that induced cells via any of the three methods were positive for TH,DAT and DAD2.Real-time RT-PCR analysis showed that TH mRNA, DAT mRNA, DRD2 mRNA expression could be seen in both induced and control group. Conclusions In this study,HUCB-MSCs is selected as the research object.By HUCB-MSCs'culture and passage,cryopreservation, recovery, HUCB-MSCs is proved that it can be stablely cultured and amplified in vitro.HUCB-MSCs can differentiate into neuron-like cells and dopaminerg- ic neuron-like cells in vitro by different methods and express dopaminergic neuron specific marker.As it indicates that their capacity can differentiate into a neruophenotype and reveals their potential use for neural transplantation.