Influence of co-culturing muscle satellite cells with preadipocytes on the differentiation of adipocytes and muscle cells isolated from Korean native cattle

C. Choi
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Abstract

The present study was done to investigate the effect of co-culturing muscle satellite cells (MSCs) and intramuscular preadipocytes (IPs) on the differentiation of adipocytes and muscle cells isolated from Korean native cattle. MSCs and IPs were single-cultured in 10% fetal bovine serum/Dulbecco's modified Eagles medium (FBS/DMEM) for 48 h followed by culturing in 5% FBS/DMEM as the growth media. Then, the growth media was replaced by differentiation media composed of 2% FBS/DMEM without any additives for the singleor co-culture of muscle cells and intramuscular adipocytes to induce the differentiation of both cell types. Cell differentiation was measured by morphological investigation and cytosolic enzyme analysis of glycerol-3-phosphate dehydrogenase (GPDH) for the adipocytes and creatine kinase (CK) for the muscle cells. In the morphological test, the presence of muscle cells did not stimulate adipocyte differentiation showing more differentiation of the adipocytes in the single-culture compared to the co-culture condition. However, the differentiation of muscle cells was promoted by adipocytes in the co-culture. The results of the enzymatic analysis were highly associated with the morphological results with a statistically higher GPDH activity (p < 0.05) appearing in the single-culture than in the co-culture, whereas the opposite was true for the CK activity of the muscle cells (p < 0.05). By manipulating in vivo the milieu using a co-culture, we could detect the difference in the rate of cell differentiation and suggest that a co-culture system is a more reliable and precise technique compared to a single-culture. Further studies on various co-culture trials including supplementation of differentiating substances, gene expression analysis, etc. should be done to obtain practical and fundamental data.
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肌肉卫星细胞与前脂肪细胞共培养对韩国土牛脂肪细胞和肌肉细胞分化的影响
本文研究了肌卫星细胞(MSCs)与肌内前脂肪细胞(IPs)共培养对牛脂肪细胞和肌肉细胞分化的影响。MSCs和IPs分别在10%胎牛血清/Dulbecco's modified Eagles培养基(FBS/DMEM)中单独培养48 h,然后在5% FBS/DMEM培养基中培养。然后用不添加任何添加剂的2% FBS/DMEM组成的分化培养基代替生长培养基,对肌细胞和肌内脂肪细胞进行单次或共培养,诱导两种细胞的分化。通过形态学观察和脂肪细胞的甘油-3-磷酸脱氢酶(GPDH)和肌肉细胞的肌酸激酶(CK)的胞质酶分析来观察细胞分化。在形态学测试中,肌肉细胞的存在并没有刺激脂肪细胞的分化,在单一培养条件下,脂肪细胞的分化比共培养条件下更多。然而,在共培养中,脂肪细胞促进了肌肉细胞的分化。酶分析结果与形态学结果高度相关,单培养GPDH活性显著高于共培养(p < 0.05),而肌细胞CK活性则相反(p < 0.05)。通过使用共培养来操纵体内环境,我们可以检测细胞分化率的差异,并表明共培养系统与单一培养相比是一种更可靠和精确的技术。进一步研究各种共培养试验,包括补充分化物质、基因表达分析等,以获得实用的基础数据。
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