Lee Youn-Kyoung, Cowan Chad A
Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA; Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.
Methods Enzymol. 2014;538:35-47. doi: 10.1016/B978-0-12-800280-3.00003-7.
Given the rapid increase in the prevalence of obesity and related metabolic diseases, research to better understand adipose tissue biology and physiology has garnered considerable attention. Adipose has been studied using both cell culture systems and model organisms. However, the mechanisms of adipocyte regulation are not comprehensively understood, as currently available in vitro or in vivo systems do not fully recapitulate human metabolic processes. Human primary adipocytes are difficult to culture and expand, and current cell systems have limitations such as cell line-to-cell line variability for adipocyte differentiation, decreased proliferation, and differentiation potential upon continued passaging. To overcome these limitations, we developed and established an efficient and robust adipocyte differentiation protocol using human pluripotent stem cells (hPSCs) and inducible expression of key adipogenic transcriptional regulators. Here, we provide a simple and stepwise protocol for programming hPSCs into mature white or brown adipocytes.
鉴于肥胖症及相关代谢疾病的患病率迅速上升,旨在更好地理解脂肪组织生物学和生理学的研究已备受关注。人们已使用细胞培养系统和模式生物对脂肪进行了研究。然而,由于目前可用的体外或体内系统无法完全重现人类代谢过程,脂肪细胞调节机制尚未得到全面了解。人原代脂肪细胞难以培养和扩增,当前的细胞系统存在诸多局限性,如脂肪细胞分化在细胞系之间存在差异、增殖能力下降以及传代后分化潜能降低。为克服这些局限性,我们利用人类多能干细胞(hPSC)和关键脂肪生成转录调节因子的诱导表达,开发并建立了一种高效且稳健的脂肪细胞分化方案。在此,我们提供了一个将hPSC编程为成熟白色或棕色脂肪细胞的简单且逐步的方案。