Technical note: Isolation and characterization of ovine brown adipocyte precursor cells
Technical note: Isolation and characterization of ovine brown adipocyte precursor cells
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技术说明：绵羊棕色脂肪细胞前体细胞的分离和表征
DOI：
10.2527/jas.2014-8728
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发表时间：
2015-05-01
影响因子：
3.3
通讯作者：
Wu, G.
中科院分区：
文献类型：
--
作者：
Ma, X.;Hou, Y. Q.;Wu, G.
棕色脂肪组织（BAT）在新生羔羊体温调节中起着重要作用。稳定的BAT细胞系的可用性对于生化研究非常宝贵，可以阐明负责胎儿BAT生长和发育营养调节的细胞和分子机制。从妊娠90 d的胎羊中分离绵羊棕色脂肪细胞前体细胞（BAPC），并进行培养，建立稳定的细胞系。这些细胞的特征在于成脂分化和标志基因解偶联蛋白1（UCP 1）的表达。BAPC每24小时增加一倍。在无血清Dulbecco改良Eagle Ham/ F12培养基中诱导9 d后，BAPC分化为棕色脂肪细胞，脂肪细胞内有较大的脂滴。分化培养基诱导BAPC表达UCP 1 mRNA和蛋白。此外，BAPC传代30次后，它们保持相似的细胞形态、成脂分化潜力和表达UCP 1的能力。综上所述，我们建立了一个稳定的绵羊BAPC细胞系，用于研究胎儿BAT生长发育的营养调控。
Brown adipose tissue (BAT) plays a critical role in regulating body temperature in newborn lambs. Availability of a stable BAT cell line would be invaluable for biochemical studies to elucidate cellular and molecular mechanisms responsible for nutritional regulation of fetal BAT growth and development. Ovine brown adipocyte precursor cells (BAPC) were isolated from fetal lambs at d 90 of gestation and cultured to establish a stable cell line. These cells were characterized by adipogenic differentiation and expression of a hallmark gene, uncoupling protein 1 (UCP1). The BAPC doubled every 24 h. After a 9-d induction with a serum-free Dulbecco's modified Eagle Ham/ F12 medium, BAPC differentiated into brown adipocytes with large lipid droplets. The differentiation medium induced expression of UCP1 mRNA and protein in BAPC. Furthermore, after BAPC were passaged 30 times, they maintained similar cell morphology, the potential for adipogenic differentiation, and the ability to express UCP1. Taken together, we have established a stable ovine BAPC cell line for studying nutritional regulation of BAT growth and development in the fetus.