Human umbilical cord blood derived mesenchymal stem cells were differentiated into pancreatic endocrine cell by Pdx-1 electrotransfer
Phuoc Thi-My Nguyen, Anh Thai-Quynh Nguyen, Nhung Thi Nguyen, Nguyet Thi-Minh Nguyen, Thu Thi Duong, Nhung Hai Truong, and Ngoc Kim Phan.
Abstract
Diabetes mellitus type 1 is an autoimmune disease with high incidence in adolescents and young adults. A seductive approach overcomes normally obstacles treatment is cell-replacement therapy to endogenous insulin production. At the present, to get enough pancreatic endocrine cells (PECs) in cell transplantation, differentiation of mesenchymal stem cells (MSCs) into IPCs is an interesting and promising strategy. This study aimed to orient umbilical cord blood-derived MSCs (UCB-MSCs) to PECs by Pdx-1 electrotransfer. UCB-MSCs were isolated from human umbilical cord blood according to published protocol. Pdx-1 was isolated and cloned into a plasmid vector. Optimal voltage of an electrotransfer was investigated to improve the cell viability and gene transfection efficacy. The results showed that 200V of the electrotransfer significantly increased in the efficiency of electrotransfer and survival cells compared with other high voltages (350V and 550V). Pdx-1 successfully transfected UCB-MSCs over-expressed pancreatic related genes as Ngn3, Nkx6.1. These results suggested that Pdx-1 transfected UCB-MSCs were successfully oriented PECs. Different to lentiviral vectors, electrotransfer is a safer method to transfer Pdx-1 to UCB-MSCs and a useful tool in translational research.
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