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Differentiation of Human Umbilical Cord-derived Mesenchymal Stem Cells to Melanocyte, Osteogenic and Neural Cells In Vitro

Yanru Zhang

This study aimed to isolated and culture human Umbilial cord Mesenchymal Stem Cells (hUMSCs) in vitro by tissue block attachment to investigate their biological characteristics. The hUMSCs were cultured to passage 3, following which the induction experiments were performed in vitro and induced to melanocytes, neurons, and osteoblasts. The Reverse Transcription-Polymerase Chain Reaction (RT-PCR) results revealed that the expression of Nanog Homeobox (NANOG), Pou Class 5 Homeobox 1 (OCT4), 5'-nucleotidase ecto (CD73), CD44 molecule (CD44) in umbilical cord-derived mesenchymal stem cells were positive and population doubling time was 24.7 hrs. The differentiation properties of neurogenesis and osteogenic cells were confirmed by histological staining using toluidine blue and Alizarin red. Melanocytes were detected by RT-PCR, quantitative PCR (qPCR), and immunofluorescence staining. After 28 days of differentiation, the cells exhibited a typical morphology such as bipolar or tripolar cells with slender dendrites. The immunofluorescence staining showed that the differentiated cells expressed Microphthalmia Transcription Factor (MITF) and Tyrosinase (TYR), the expression of MITF, TYR and KIT proto-oncogene receptor tyrosine Kinase (KIT) from 0 d to 28 days by qPCR was significantly different and the expression of marker genes MITF, SOX10, KIT by RT-PCR were positive. The results demonstrated that the cells isolated from human umbilical cord were mesenchymal stem cells, and hUMSCs had multidifferentiation potentialities. hUMSCs could also differentiate into melanocytes in vitro, providing reliable sources of melanocytes for treating vitiligo future.