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Differentiation of porcine adipose mesenchymal stem cells into adipocytes and the expression of Krüppel like factor 2
LIU Jing-xia LI Fang-zheng PIAO Ying-ji JIANG Zhong-ling SONG Xue-xiong*
Acta Anatomica Sinica ›› 2015, Vol. 46 ›› Issue (5) : 616-622.
Differentiation of porcine adipose mesenchymal stem cells into adipocytes and the expression of Krüppel like factor 2
Objective To investigate the gene expression changes of Krüppe-1 like factor (KLF)2 during the process of induced adipogenic differentiation from porcine adipose mesenchymal stem cells(AMSCs) in vitro and to reveal the foundation for regulating the differentiation of adipocyte. Methods The porcine AMSCs were isolated by typeⅠcollagen enzyme method and subcultured. F3 culture cell surface antigen was detected by flow cytometry. The differentiated adipocytes were identified by morphology and oil red O staining; The expression of KLF2 mRNA and adipogenic marker genes proliferator-activated receptor γ(PPARγ)2 were measured by Real-time PCR. Results The AMSCs expressed the MSCs cellular markers, CD29,CD44 and CD105 with a marking rate a of 91.7%,95.1% and 95.6% respectively,but did not express CD34 with a marking rate of 3.39%. Some adipocytes appeared when the AMSCs induced and differentiated for 2 days. When the induced differentiation time extended, the number of adipocytes increased. The adipogenic conversion rate reached 48.2% in the induction of 18 days. In the induction of 2,8 and 16 days, the expressions of KLF2 mRNA were 1.84±0206,1.07±0.072 and 0.83±0.095,and the expressions of PPARγ2 mRNA were 3.06±0.542,13.22±0.5736 and 15.11±1.073. Conclusion This study successfully obtained AMSCs which showed good adipogenic differentiation potential.The expressions of KLF2 was increased in early adipogenic differentiation phase, and then gradually reduced.The results suggest that the KLF2 is the negative regulation of transcription factors.
Adipose mesenchymal stem cell / Surface antigen / Induction / Differentiation / Krüppe-1 like factor 2 / Real-time PCR / Porcine
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