Correlation of spontaneous adipocyte generation with osteogenic differentiation of porcine skin-derived stem cells
- Author:
Hyo Kyung BAE
1
;
Bae Dong JUNG
;
Seunghyung LEE
;
Choon Keun PARK
;
Boo Keun YANG
;
Hee Tae CHEONG
Author Information
- Publication Type:Original Article
- Keywords: Porcine skin-derived stem cells; Osteogenesis; Spontaneous adipocyte generation; Peroxisome proliferator-activated receptor gamma 2
- MeSH: Adipocytes; Alkaline Phosphatase; Calcium; Cell Line; Ether; In Vitro Techniques; Lipid Droplets; Osteocytes; Osteogenesis; PPAR gamma; RNA, Messenger; Stem Cells; Transcription Factors
- From:Journal of Veterinary Science 2019;20(1):16-26
- CountryRepublic of Korea
- Language:English
- Abstract: The objective of this study was to examine effects of spontaneous adipocyte generation on osteogenic differentiation of porcine skin-derived stem cells (pSSCs). Correlation between osteogenic differentiation and adipocyte differentiation induced by osteocyte induction culture was determined using different cell lines. Osteogenic differentiation efficiency of pSSCs was then analyzed by controlling the expression of adipocyte-specific transcription factors during osteogenic induction culture. Among four cell lines, pSSCs-II had the lowest lipid droplet level but the highest calcium content (p < 0.05). It also expressed significantly low levels of peroxisome proliferator-activated receptor gamma 2 (PPARγ2) and adipocyte protein 2 (aP2) mRNAs but very high levels of runt-related transcription factor 2 (Runx2) and alkaline phosphatase (ALP) mRNAs as osteogenic makers (p < 0.05). Oil red O extraction was increased by 0.1 µM troglitazone (TGZ) treatment but decreased by 50 µM bisphenol A diglycidyl ether (BADGE) (p < 0.05). Calcium content was drastically increased after BADGE treatment compared to that in osteogenic induction control and TGZ-treated pSSCs (p < 0.05). Relative expression levels of PPARγ2 and aP2 mRNAs were increased by TGZ but decreased by BADGE. Expression levels of Rucx2 and ALP mRNAs, osteoblast-specific marker genes, were significantly increased by BADGE treatment (p < 0.05). The expression level of BCL2 like 1 was significantly higher in BADGE-treated pSSCs than that in TGZ-treated ones (p < 0.05). The results demonstrate that spontaneous adipocyte generation does not adversely affect osteogenic differentiation. However, reducing spontaneous adipocyte generation by inhibiting PPARγ2 mRNA expression can enhance in vitro osteogenic differentiation of pSSCs.