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

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 mM cortisol and 2 mM BRL 49653, a PPARg 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-speci®c 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

Functional expression of glucose-dependent insulinotropic polypeptide receptors is coupled to differentiation in a human adipocyte model

“…15 A number of studies have established that the pattern and time-course of gene expression in SGBS cells during differentiation is comparable to the findings in human preadipocytes in primary culture and also resemble the characteristics of rodent preadipocyte cell lines such as 3T3-L1, 3T3-F442A and ob17. 15,16,[20][21][22][23][24] Differentiated SGBS cells also behave biochemically and functionally like human adipocytes differentiated in primary culture. 15,16,25 In this study, we used these cells to examine the regulation of GIPR expression in the context of the dynamic process of adipocyte differentiation.…”

“…15,16,[20][21][22][23][24] Differentiated SGBS cells also behave biochemically and functionally like human adipocytes differentiated in primary culture. 15,16,25 In this study, we used these cells to examine the regulation of GIPR expression in the context of the dynamic process of adipocyte differentiation. Human adipocytes express functional GIP receptors RE Weaver et al…”

“…For this purpose, we used the recently established in vitro model of Simpson-GolabiBehmel syndrome (SGBS) human preadipocytes, a cell strain, which has the capacity to differentiate into mature adipocytes under defined experimental conditions. 15 …”

4-hydroperoxy-cyclophosphamide mediates caspase-independent T-cell apoptosis involving oxidative stress-induced nuclear relocation of mitochondrial apoptogenic factors AIF and EndoG

“…This was further confirmed in a second cellular system using the preadipocyte cell line SGBS. 30 4-OOH-CY-induced apoptosis in SGBS was also largely caspase-independent since z-VAD.fmk did not efficiently block apoptosis induction, while NAC totally prevented cell death (Supplementary Figure 3). Inhibition of EndoG (Figure 7c) and AIF (Figure 7d) expression in SGBS cells efficiently protected from 4-OOH-CY-mediated apoptosis.…”

Vaspin is related to gender, puberty and deteriorating insulin sensitivity in children

“…In addition, we applied pre-adipocytes and adipocytes of the human SGBS cell line. 15 Reverse transcription of RNAs was performed using 200 U M-MLV reverse transcriptase per microgram of total RNA (Invitrogen, Karlsruhe, Germany) with oligo (dT) primers. Vaspin mRNA expression was determined using real-time PCR with TaqMan probe-based gene expression assay on the ABI 7500 Sequence Detection System (Applied Biosystems, Darmstadt, Germany).…”