Isolation of a preadipocyte cell line from rat bone marrow and differentiation to adipocytes.
Isolation of a preadipocyte cell line from rat bone marrow and differentiation to adipocytes.
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DOI：
10.1210/endo.136.10.7545105
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发表时间：
1995-10
期刊：
影响因子：
4.8
通讯作者：
O. Marko;M. Cascieri;N. Ayad;C. Strader;M. Candelore
中科院分区：
文献类型：
--
作者：
O. Marko;M. Cascieri;N. Ayad;C. Strader;M. Candelore
一组独特的大鼠脂肪细胞前体细胞来源于正常大鼠骨髓。采用差速胰酶消化、Ficoll梯度离心法和差速接种相结合的方法，从混合培养的骨髓细胞中分离出上皮样前脂肪细胞。该细胞系命名为RBM-Ad，可完全分化为形态类似棕色脂肪组织的多房脂肪细胞。在这些细胞的增殖过程中，没有观察到分化模式的变化，它们已经成功地携带和分化到第49代。苏丹黑、苏丹IV和油红O对分化细胞的组织学染色表明，细胞内小泡中存在脂质。非选择性β-肾上腺素能激动剂异丙肾上腺素刺激前脂肪细胞和分化脂肪细胞的腺酰环化酶活性。相比之下，β3-肾上腺素能受体特异性激动剂BRL-37344能刺激分化脂肪细胞的腺酰环化酶活性和甘油释放，但不能刺激前脂肪细胞。此外，分化后的脂肪细胞含有信使RNA，编码棕色脂肪特有的蛋白质--热原。因此，该大鼠前脂肪细胞系可分化为在组织结构和功能上与棕色脂肪组织相似的脂肪细胞。
A unique population of rat adipocyte precursor cells was derived from normal rat bone marrow. The epitheloid-like preadipocytes were isolated from a mixed culture of bone marrow cells by a combination of differential trypsinization, enrichment by Ficoll gradient centrifugation, and differential seeding. This cell line, designated RBM-Ad, can be fully differentiated into multilocular adipocytes morphologically resembling brown adipose tissue. No changes in the differentiation pattern are observed during propagation of these cells, and they have been successfully carried and differentiated up to passage 49. Histological staining of differentiated cells with Sudan black, Sudan IV, and oil red O indicates the presence of lipids in intracellular vesicles. The nonselective beta-adrenergic agonist isoproterenol stimulates adenylyl cyclase activity in both preadipocytes and differentiated adipocytes. In contrast, BRL-37344, a beta 3-adrenergic receptor-specific agonist, stimulates adenylyl cyclase activity and glycerol release in differentiated adipocytes, but not preadipocytes. In addition, differentiated adipocytes contain messenger RNA encoding the brown adipose-specific protein, thermogenin. Thus, this rat preadipocyte cell line can be differentiated into adipocytes that histologically and functionally resemble brown adipose tissue.