Bohan Amy E, Purvis Katelyn N, Bartosh Julia L, Brandebourg Terry D
Department of Animal Sciences; Auburn University ; Auburn, AL USA.
Adipocyte. 2014 Dec 10;3(4):322-32. doi: 10.4161/21623945.2014.981434. eCollection 2014 Oct-Dec.
Given similarities in metabolic parameters and cardiovascular physiology, the pig is well positioned as a biomedical model for metabolic disease and obesity in humans. Better understanding molecular mechanisms governing porcine adipocyte hyperplasia may provide insight into the regulation of adipose tissue development that is useful both when considering the pig as a commodity and when extrapolating porcine data to human disease. Primary cultures of pig stromal-vascular cells have served as a useful tool for investigating factors that regulate preadipocyte proliferation and differentiation. However, such cultures have generally been maintained at 37°C in vitro despite euthermia being 39°C in pigs. To address potential concerns about the physiological relevance of culturing primary pig preadipocytes under what would be hypothermic conditions in vivo, the objective of this study was to investigate the effect of culture temperature on the proliferation and differentiation of pig preadipocytes in primary culture. Culturing primary preadipocytes at 37 rather than 39°C decreases their proliferation rates based upon cleavage of the tetrazolium salt, MTT (P < 0.001), reduction of resazurin (P < 0.001), and daily cell counts (P < 0.001). Likewise, culturing primary porcine preadipocytes at 37°C suppressed their adipogenic potential based upon monitoring adipogenesis morphologically, biochemically, and via the expression of mRNA encoding adipogenic marker genes. Collectively, these data indicate the proliferation and differentiation of primary pig preadipocytes is suppressed when cultures are incubated at 37°C compared to normal body temperature of pigs. This may confound investigation of factors that impact adipocyte hyperplasia in the pig.
鉴于猪在代谢参数和心血管生理学方面与人类存在相似性,猪是研究人类代谢疾病和肥胖症的理想生物医学模型。深入了解控制猪脂肪细胞增生的分子机制,有助于洞察脂肪组织发育的调控机制,这对于将猪作为商品来考量以及将猪的数据外推至人类疾病研究都具有重要意义。猪基质血管细胞的原代培养,已成为研究调节前脂肪细胞增殖和分化因素的有效工具。然而,尽管猪的正常体温为39°C,但此类培养通常在体外37°C条件下进行。为解决在体内低温条件下培养猪原代前脂肪细胞可能存在的生理相关性问题,本研究旨在探讨培养温度对原代培养的猪前脂肪细胞增殖和分化的影响。基于四氮唑盐MTT的裂解(P < 0.001)、刃天青的还原(P < 0.001)以及每日细胞计数(P < 0.001),在37°C而非39°C培养原代前脂肪细胞会降低其增殖速率。同样,基于形态学、生化监测以及脂肪生成标记基因mRNA表达情况,在37°C培养原代猪前脂肪细胞会抑制其脂肪生成潜能。总体而言,这些数据表明,与猪的正常体温相比,在37°C培养时,原代猪前脂肪细胞的增殖和分化受到抑制。这可能会干扰对影响猪脂肪细胞增生因素的研究。