Establishment Of Somatic Partially Reprogrammed Cells And Its Plasticity Research

Induced pluripotent technology has become important experimental technology for cell proliferation, differentiation, development and regenerative medicine researches. At present, innovation of induced pluripotent technology, mechanism of somatic reprogramming, screening for therapeutic compounds and regeneration medicine are important research field in induced pluripotent stem cells (iPSCs). With the deepening of somatic reprogramming, researchers inferred that there maybe a reprogramming intermediates during reprogramming. Recently, epigenetic modifier, Aicda, key factor of Mesenchymal-to-Epithelial Transition, E-cadherin and maternal gene in mammal oocyts, Glis1were used to replace traditional transcriptional factor (Klf4, Sox2, c-Myc and Oct4), and estimate the effect of these factors for establishing reprogramming intermediates. In this study, C57-EGFP fibroblasts were co-transduced with Klf4, Sox2, c-Myc and Oct4lentivirus, and C57-EGFP-4F iPSCs were achieved. The cells were most like mouse embryonic stem cells in morphology, pluripotent mark genes expression, embryoid body formation and generation of chimaeric mouse. Using this transduction system, a transcriptional factor cocktail with Aicda, E-cadherin, Glis1, Klf4, Sox2, c-Myc and Oct4was used to produce iPSCs. However, cocktail with Aicda was significantly decrease stem cells like colonies formation during reprogramming. When removing Aicda, stem like cells were observed by transducing Klf4, c-Myc, E-cadherin and Glis1transductiong. Analysized cells character showed that the cells did not express AP, Oct4, Sox2and Nanog, but SSEA-1positive. When culturing them into petri-dishes, they formed embryoid body, and can only differentiate into ectoderm (nestin positive) and mesoderm (fltl positive) compare with ESCs and iPSCs. So, we called it KMGE partially reprogrammed cells (KMGE-prCs). Subsequently, for detecting its secondary reprogramming, KMGE-prCs were treated with small molecular compounds, PD0325901, CHIR99021and A-83-01. The results showed that there were no significant change of morphology after treatment, and the pluripotent marker genes expression were not reactivated. However, when formed embryoid body, KMGE-prCs differentiated into ectoderm (nefl and nestin positive), mesoderm (fltl positive) and endoderm (gata4positive). Thereforce, we get partially reprogrammed cells with specific plasticity. We will further analyze characteristics of the cells in the future study, and using it to somatic reprogramming mechanism research.