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| Optimizing Cultivation of Mature Adipocytes and Obtaining Preadipocytes by Mature Adipocyte Dedifferentiation in Pigs(Sus scrofa) |
| , , , , ,Jie CHEN |
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Abstract The mature adipocyte dedifferentiation is able to provide homogeneous preadipocyees for researching adpocyte differentiation. In this study, mature adipocytes were isolated and cultured from adipose tissue, and then dedifferentiated to achieve preadipocytes. The subcutaneous adipose tissue from 3-day-old piglets(Sus scrofa) were digested by collagenase type Ⅱ, followed by centrifugation at different centrifugal force and then cultured by "ceiling culture" method. Morphological changes of adipocytes were observed under microscope, and the degree of adipogenesis and differentiation was assessed via oil red O staining. By inducing, adipose-derived progeny cells were redifferentiated into lipid-laden cells with accumulation of lipids. In terms of expression level, peroxisome proliferator-activated receptor-γ (PPARγ) and fatty acid binding protein 4 (FABP4) were detected by Real-time PCR. As we expected, the adipogenic markers, PPARγ and FABP4, increased along with the redifferentiation process. Particularly, a 2.8-fold increase of PPARγ expression, and a 62-fold of FABP4 were shown in the late phase of the redifferentiation process, demonstrating the significantly higher expression levels comparing with the unredifferentiated cells at 0 d(P<0.05). Showing that the predipocytes obtained by dedifferentiation might effectively differentiate to mature adipocytes with adipogenic induction agent. In addition, our study optimized the mature adipocytes culture system as well. Using this system, mature adipocytes can revert into proliferative-competent progeny cells by inducing redifferentiate again into mature adipocytes, which contributes an in vitro model for adipocytes investigation
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Received: 05 July 2012
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Corresponding Authors:
Jie CHEN
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