Transdifferentiation of porcine satellite cells to adipoblasts with ciglitizone

¹ Product and Utility Division, Chonbuk National University
² Department of Animal Resources and Biotechnology, Chonbuk National University
³ Functional Animal Genomics, National Livestock Research Institute, Suwon 441-350, Republic of Korea
⁴ Nutrition and physiology, National Livestock Research Institute, Suwon 441-350, Republic of Korea
⁵ Animal Genomics and Bioinformatics Division, National Livestock Research Institute, Suwon 441-350, Republic of Korea

^* To whom correspondence should be addressed. E-mail: hs6226{at}rda.go.kr.

	Abstract

Ciglitizone, a class of thiazolidinediones (TZD) acts as potent activators of the adipose differentiation program in established preadipose cell lines. Thiazolidinediones have also been investigated in diabetic patients and reported to act as peroxisome proliferated activated receptor- gamma (PPAR-) ligands. Intramuscular adipogenesis or marbling through transdifferentiation of satellite cells in cattle has been successfully conducted earlier. This study was therefore, taken up in a different species. In this report, the effects of ciglitizone on the differentiation pathway of porcine myogenic satellite cells have been investigated. Semitendinosus muscle was aseptically taken from 10 d old piglets under general anesthesia. Connective tissues were removed and protease digestion of excised muscle was done after fine meshing. Finally, porcine satellite cells were obtained and grown to almost 100% confluence. Post confluent cells (d 0) were further cultured in differentiation medium containing Dulbecco's modified Eagle's Medium + 2% horse serum + 1% penicillin/streptomycin with adipogenic mixture plus ciglitizone (10µM) for 48 h. From d 2 onwards cells were cultured only in the presence of ciglitizone (10µM) every 48 h until d 10. Controls were cultured in differentiation medium only. Exposure of porcine satellite cells to adipogenic mixture and ciglitizone generated lipid droplets on d 2 and subsequently using ciglitizone alone helped in cytoplasmic lipid filling providing the cells with the acquisition of adipocyte morphology. Increase (P < 0.05) in the fusion (structure containing 2 to 3 nuclei) of satellite cells was observed and myosin heavy chain appeared with greater intensity (immunohistochemistry) in the control group from d 2 onwards. Adipocyte specific transcriptional factors (i.e. CCAAT/enhancer binding protein- alpha and PPAR- were predominant during transdifferentiation and were observed with immuhistochemistry, western blot (~ 47.2 and ~ 60.4 kDa respectively) and real time PCR. Ciglitizone appeared to convert the differentiation pathway of satellite cells into that of adipoblasts.

Key Words: Adipogenesis, Ciglitizone , C/EBP- myogenesis, PPAR-

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