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KMID : 1005220160310010089
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Journal of Embryo Transfer
2016 Volume.31 No. 1 p.89 ~ p.95
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Analysis of ROS and Apoptosis of Porcine Skin-derived Stem-like Cells after Differentiation Induction into Mesodermal Cell Types
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Bae Hyo-Kyung
Lee Hwa-Yeon
Park Yeo-Reum
Park Choon-Keun
Yang Boo-Keun
Cheong Hee-Tae
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Abstract
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The present study was conduct to examine the H2O2 expression level and apoptosis-related gene expression levels inporcineskin-derived stem cell-like cells (pSSCs) after adipogenic, chondrogenic, and osteogenic differentiation induction. The pSSCs were obtained by digestion of porcine ear skin biopsy and cultured in each induction medium for 21 to 26 days to induce adipogenic, chondrogenic, and osteogenic differentiation, respectively. The H2O2 levels of pSSCs after induction culture were evaluated by staining with 2'7'-dichlorodihydrofluorescein diacetate (H2DCFDA). The apoptotic gene expression of pSSCs after induction culture was also estimated by RT-PCR. The pSSCs have a potential to differentiate into three mesodermal cell types (adipocytes, chondrocytes, and osteoblasts). Non-induced control and chondrogenic-induced cells were showed higher H2DCFDA intensity (P<0.05) than adipogenic- and osteogenic-induced cells. The relative expression of Bax/Bcl-2 level was significantly low (P<0.05) in adipogenic- and osteogenic-induced cells compared to non-induced control. However, there was no difference in the relative expression of Bax/Bcl-2 level among differentiation induction groups. The result of the present study shows that the apoptosis of pSSCs is not detrimentally increased by differentiation induction culture, although chondrogenic-induced pSSCs showed high ROS generation level and apoptotic index similarly to those of non-induced cells.
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KEYWORD
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porcine skin-derived stem-like cells (pSSCs) , differentiation induction , ROS , apoptosis
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