Isolation and Differentiation of Primary White and Brown Preadipocytes from Newborn Mice.
Isolation and Differentiation of Primary White and Brown Preadipocytes from Newborn Mice.
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新生小鼠原代白色和棕色前脂肪细胞的分离与分化。
DOI：
10.3791/62005
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发表时间：
2021-01-25
期刊：
影响因子：
--
通讯作者：
Saez E
中科院分区：
文献类型：
--
作者：
Galmozzi A;Kok BP;Saez E
对脂肪细胞分化和功能机制的理解很大程度上得益于永生化白色前脂肪细胞系的使用。然而，这些培养的细胞系有局限性。他们没有完全捕捉到现在已知存在于白色脂肪库中的异质脂肪细胞群的不同功能谱。为了提供一个生理学上更相关的模型来研究白色脂肪组织的复杂性，研究人员开发并优化了一种方案，可以同时从新生小鼠中分离原代白色和棕色脂肪细胞祖细胞，使其在培养中快速扩增，并在体外分化为成熟的、功能齐全的脂肪细胞。从新生小鼠而不是成年小鼠中分离原代细胞的主要优点是，脂肪库正在积极发育，因此是增殖前脂肪细胞的丰富来源。使用该方法分离的原代前脂肪细胞在达到合流后迅速分化，并在4-5天内完全成熟，这一时间窗口准确反映了新生小鼠发育的脂肪垫的外观。使用这种策略制备的原代培养物可以扩展和研究，具有高重复性，使其适用于遗传和表型筛选，并能够研究遗传小鼠模型的细胞自主脂肪细胞表型。该方案提供了一个简单，快速，廉价的方法来研究体外脂肪组织的复杂性。
The understanding of the mechanisms underlying adipocyte differentiation and function has greatly benefited from the use of immortalized white preadipocyte cell lines. These cultured cell lines, however, have limitations. They do not fully capture the diverse functional spectrum of the heterogenous adipocyte populations that are now known to exist within white adipose depots. To provide a more physiologically relevant model to study the complexity of white adipose tissue, a protocol has been developed and optimized to enable simultaneous isolation of primary white and brown adipocyte progenitors from newborn mice, their rapid expansion in culture, and their differentiation in vitro into mature, fully functional adipocytes. The primary advantage of isolating primary cells from newborn, rather than adult mice, is that the adipose depots are actively developing and are, therefore, a rich source of proliferating preadipocytes. Primary preadipocytes isolated using this protocol differentiate rapidly upon reaching confluence and become fully mature in 4-5 days, a temporal window that accurately reflects the appearance of developed fat pads in newborn mice. Primary cultures prepared using this strategy can be expanded and studied with high reproducibility, making them suitable for genetic and phenotypic screens and enabling the study of the cell-autonomous adipocyte phenotypes of genetic mouse models. This protocol offers a simple, rapid, and inexpensive approach to study the complexity of adipose tissue in vitro.
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影响因子：
29
作者：
Kajimura S;Spiegelman BM;Seale P
通讯作者：
Seale P
克隆衍生的成年人类棕色脂肪细胞的遗传和功能表征。
影响因子：
82.9
作者：
Shinoda, Kosaku;Luijten, Ineke H. N.;Hasegawa, Yutaka;Hong, Haemin;Sonne, Si B.;Kim, Miae;Xue, Ruidan;Chondronikola, Maria;Cypess, Aaron M.;Tseng, Yu-Hua;Nedergaard, Jan;Sidossis, Labros S.;Kajimura, Shingo
通讯作者：
Kajimura, Shingo
影响因子：
5.2
作者：
Lynes MD;Tseng YH
通讯作者：
Tseng YH
影响因子：
5.8
作者：
Brown EL;Hazen BC;Eury E;Wattez JS;Gantner ML;Albert V;Chau S;Sanchez-Alavez M;Conti B;Kralli A
通讯作者：
Kralli A
影响因子：
64.8
作者：
Galmozzi, Andrea;Kok, Bernard P.;Saez, Enrique
通讯作者：
Saez, Enrique