一种具有高脂肪分化能力的人前脂肪细胞系的特性研究

Characterization of a human preadipocyte cell strain with high capacity for adipose differentiation.

作者信息

Wabitsch M, Brenner R E, Melzner I, Braun M, Möller P, Heinze E, Debatin K M, Hauner H

机构信息

Department of Pediatrics, University of Ulm, Ulm, Germany.

出版信息

Int J Obes Relat Metab Disord. 2001 Jan;25(1):8-15. doi: 10.1038/sj.ijo.0801520.

DOI:10.1038/sj.ijo.0801520

PMID:11244452

Abstract

OBJECTIVE

To develop and to characterize a human preadipocyte cell strain with high capacity for adipose differentiation serving as a model for studying human adipocyte development and metabolism in vitro.

METHODS

Cells were derived from the stromal cells fraction of subcutaneous adipose tissue of an infant with Simpson-Golabi-Behmel syndrome (SGBS). Adipose differentiation was induced under serum-free culture conditions by exposure to 10 nM insulin, 200 pM triiodothyronine, 1 microM cortisol and 2 microM BRL 49653, a PPAR gamma agonist.

RESULTS

During the differentiation process SGBS cells developed a gene expression pattern similar to that found in differentiating human preadipocytes with a characteristic increase in fat cell-specific mRNAs encoding lipoprotein lipase (LPL), glycero-3-phosphate dehydrogenase (GPDH), GLUT4, leptin and others. Differentiated SGBS cells exhibited an increase in glucose uptake upon insulin stimulation and in glycerol release upon catecholamine exposure. SGBS adipocytes were morphologically, biochemically and functionally identical to in vitro differentiated adipocytes from healthy subjects. However, while preadipocytes from healthy control infants rapidly lost their capacity to differentiate after a few cell divisions in culture, SGBS cells maintained their differentiation capacity over many generations: upon appropriate stimulation 95% of SGBS cells of generation 30 developed into adipocytes. A mutation in the glypican 3 gene was not detected in the patient. Thus, it remains unclear whether the molecular alteration in SGBS cells is also responsible for the high differentiation capacity and further investigations are required.

CONCLUSION

The human cell strain described here provides an almost unlimited source of human preadipocytes with high capacity for adipose differentiation and may, therefore, represent a unique tool for studying human fat cell development and metabolism. International Journal of Obesity (2001) 25, 8-15

摘要

目的

建立并鉴定一种具有高脂肪分化能力的人前脂肪细胞系，作为体外研究人脂肪细胞发育和代谢的模型。

方法

细胞来源于一名患有辛普森-戈拉比-贝赫梅尔综合征（SGBS）婴儿的皮下脂肪组织的基质细胞部分。在无血清培养条件下，通过暴露于10 nM胰岛素、200 pM三碘甲状腺原氨酸、1 μM皮质醇和2 μM BRL 49653（一种PPARγ激动剂）来诱导脂肪分化。

结果

在分化过程中，SGBS细胞形成了一种与分化中的人前脂肪细胞相似的基因表达模式，编码脂蛋白脂肪酶（LPL）、甘油-3-磷酸脱氢酶（GPDH）、GLUT4、瘦素等的脂肪细胞特异性mRNA有特征性增加。分化后的SGBS细胞在胰岛素刺激下葡萄糖摄取增加，在儿茶酚胺暴露下甘油释放增加。SGBS脂肪细胞在形态、生化和功能上与健康受试者体外分化的脂肪细胞相同。然而，健康对照婴儿的前脂肪细胞在培养中经过几次细胞分裂后迅速丧失分化能力，而SGBS细胞在许多代中都保持了分化能力：在适当刺激下，第30代的95%的SGBS细胞发育成脂肪细胞。在患者中未检测到磷脂酰肌醇蛋白聚糖3基因的突变。因此，尚不清楚SGBS细胞中的分子改变是否也导致了高分化能力，需要进一步研究。