猪DFAT细胞成脂再分化过程中KLF2和PPARγ的表达

doi:10.11843/j.issn.0366-6964.2017.01.008

Abstract

Abstract:

To investigate the expression patterns of Kruppel like factor 2 (KLF2) and peroxisome proliferator activated receptor gamma (PPARγ) during the adipogenic redifferentiation of porcine dedifferentiated fat (DFAT) cells and lay a foundation for revealing the mechanism of porcine DFAT cell redifferentiation, we applied the “ceiling” culture to obtain porcine DFAT cells from mature adipocytes, detected the surface antigens of DFAT cells by flow cytometry, and then, identified the adipogenic differentiation of DFAT cells by methods of morphology, oil red O staining and real-time PCR. The results showed that the dedifferentiation sign was clearly observed, showing that the horn like protrusions were formed, large lipid droplets were decomposed into small lipid droplets, and lipid droplets were extracellularly discharged after porcine mature adipocytes were seeded on the third day, the lipid droplets were almost discharged in 9 day, and the bottom wall was covered with primary cells in 12 day. The positive rates of mesenchymal stem cell surface antigen CD29, CD44 and CD105 were 99.0%, 97.8% and 99.5%, respectively, while the percentages of hematopoietic stem cell surface antigen CD45 and CD34 were only 3.55% and 4.55%. Then, after 3 days of adipogenic induction of F3 generation DFAT cells, the spindle shaped cells became wider and shorter, and lipid droplets were observed in the cytoplasm. With the increase of induction time, the number and volume of lipid droplets increased, and the induction efficiency was more than 80% after 12 day induction. During the progress of adipogenic induction, the relative expression levels of KLF2 were 0.47±0.02, 0.35±0.07, 0.31±0.09 and 0.11±0.07, and PPARγ expression levels were 1.62±0.01, 2.03±0.04, 3.22±0.04 and 3.49±0.06 on the 2, 5, 10 and 15 days, respectively. In conclusion, porcine DFAT cells with high purity were successfully obtained, owning the mesenchymal stem cell properties and the adipogenic redifferentiation competence, and, KLF2 expression levels decreased, while PPARγ expression increased with the extension of induction time during adipogenic redifferentiation of DFAT cells, indicating that KLF2 might play an inhibitory role, while PPARγ could take on a positive role during the adipogenic redifferentiation of DFAT cells.

CLC Number:

S828.2

GAO Xia, LI Fang-zheng, HUAN Yan-jun, LIU Zhi-min, LI Xiu, JIANG Zhong-ling, NIU De-xun, SONG Xue-xiong. The Expression Patterns of KLF2 and PPARγ during the Adipogenic Redifferentiation of Porcine DFAT Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(1): 68-74.

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The Expression Patterns of KLF2 and PPARγ during the Adipogenic Redifferentiation of Porcine DFAT Cells

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