The extract of pluripotent stem cells induces dedifferentiation of somatic cells with restricted plasticity. In this study, we used the extract of human embryonic stem cells (hESC) to dedifferentiate adipose tissue-derived stem cells (ADSCs) and examined the impact of this reprogramming event on dopaminergic differentiation of the cells. For this purpose, cytoplasmic extract of ESCs was prepared by repeated cycles of freezing and thawing. The plasma membrane of hADSCs was reversibly permeabilized by Streptolysin O (SLO), exposed to hESC extract and resealed by CaCl2-containing medium. As revealed by qPCR analysis, expression of OCT4, SOX2, NANOG, LIN28A and KLF4 mRNAs were downregulated in the ADSCs one week after extract incubation, while all except for KLF4 were upregulated at the end of second week. For dopaminergic differentiation, control and reprogrammed ADSCs were induced by serum-free neurobasal medium containing B27 and a cocktail of SHH, FGF8, bFGF and BDNF for 12 days. After differentiation, expression levels of some neuronal and dopaminergic-related genes, including PAX6, NESTIN, NEFL, GLI1, LMXB1, EN1, NURR1 and TH, were significantly increase in the reprogrammed ADSCs compared to the control group. As a whole, two weeks after reprogramming by ESC extract, ADSCs showed an improved dopaminergic differentiation potential. These findings suggest that the cytoplasmic extract of hESCs is containing some regulatory factors which induce the expression of pluripotency-associated markers in somatic cells and that the exposure to ESC extract may serve as a simple and rapid strategy to enhance plasticity of somaticstem cells for cell replacement therapy purposes.
Human embryonic stem cells which express hrGFP in the undifferentiated state and during dopaminergic differentiation