Scaffold-free generation of uniform adipose spheroids for metabolism research and drug discovery.
Scaffold-free generation of uniform adipose spheroids for metabolism research and drug discovery.
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DOI：
10.1038/s41598-017-19024-z
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发表时间：
2018-01-11
影响因子：
4.6
通讯作者：
Ankrum JA
中科院分区：
文献类型：
--
作者：
Klingelhutz AJ;Gourronc FA;Chaly A;Wadkins DA;Burand AJ;Markan KR;Idiga SO;Wu M;Potthoff MJ;Ankrum JA
脂肪组织功能障碍对II型糖尿病和其他代谢疾病的发展至关重要。虽然单层细胞培养在研究脂肪生物学方面很有用，但二维培养往往不能反映脂肪组织的复杂性。动物模型也存在问题，因为它们昂贵、耗时，而且由于物种变异，可能无法完全概括人类生物学。为了解决这些问题，我们开发了一种无支架的方法，从原代或不朽的人或小鼠前脂肪细胞中生成三维脂肪球体。前脂肪细胞在悬挂滴液中自组织成球体，并转移到低附着板上，可在长期培养中维持。在暴露于分化线索后，细胞成熟为脂肪细胞，积累大的脂滴，随着时间的推移而扩大。与2D培养相比，3D球体表达和分泌更高水平的脂联素，并通过分泌促炎脂肪因子来应对培养相关或毒素相关的压力。此外，来自棕色脂肪组织（BAT）的三维球体比来自相同组织的二维培养物更好地保留了BAT标记的表达。因此，该模型既可以用于研究预脂肪细胞的成熟，也可以用于研究成熟脂肪细胞在三维培养环境中的功能。
Adipose tissue dysfunction is critical to the development of type II diabetes and other metabolic diseases. While monolayer cell culture has been useful for studying fat biology, 2D culture often does not reflect the complexity of fat tissue. Animal models are also problematic in that they are expensive, time consuming, and may not completely recapitulate human biology because of species variation. To address these problems, we have developed a scaffold-free method to generate 3D adipose spheroids from primary or immortal human or mouse pre-adipocytes. Pre-adipocytes self-organize into spheroids in hanging drops and upon transfer to low attachment plates, can be maintained in long-term cultures. Upon exposure to differentiation cues, the cells mature into adipocytes, accumulating large lipid droplets that expand with time. The 3D spheroids express and secrete higher levels of adiponectin compared to 2D culture and respond to stress, either culture-related or toxin-associated, by secreting pro-inflammatory adipokines. In addition, 3D spheroids derived from brown adipose tissue (BAT) retain expression of BAT markers better than 2D cultures derived from the same tissue. Thus, this model can be used to study both the maturation of pre-adipocytes or the function of mature adipocytes in a 3D culture environment.
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影响因子：
5.2
作者：
Kwon H;Pessin JE
通讯作者：
Pessin JE
影响因子：
29
作者：
Berry R;Jeffery E;Rodeheffer MS
通讯作者：
Rodeheffer MS
影响因子：
10
作者：
Abbott RD;Wang RY;Reagan MR;Chen Y;Borowsky FE;Zieba A;Marra KG;Rubin JP;Ghobrial IM;Kaplan DL
通讯作者：
Kaplan DL
影响因子：
3.3
作者：
Prajapati, Bhumika;Jena, Prasant Kumar;Seshadri, Sriram
通讯作者：
Seshadri, Sriram
DOI：
10.1073/pnas.1008117107
发表时间：
2010-08-03
影响因子：
11.1
作者：
Bartosh, Thomas J.;Ylostalo, Joni H.;Prockop, Darwin J.
通讯作者：
Prockop, Darwin J.