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

International Journal of Obesity volume 25, pages 8–15 (2001)Cite this article

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 µM cortisol and 2 µM BRL 49653, a PPARγ 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.

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References

  1. Spiegelman BM, Flier JS . Adipogenesis and obesity: rounding up the big picture Cell 1996 87: 377–389.

    Article  CAS  Google Scholar 

  2. Ailhaud G, Hauner H . Development of white adipose tissue. In: Bray G, Bouchard C, James WPT (eds) Handbook of Obesity Marcel Dekker: New York 1997 pp 359–378.

    Google Scholar 

  3. Zilberfarb V, Pietri-Rouxel F, Jockers R, Krief S, Delouis C, Issad T, Strosberg, AD . Human immortalized brown adipocytes express functional beta3-adrenoreceptor coupled to lipolysis J Cell Sci 1997 110: 801–807.

    CAS  PubMed  Google Scholar 

  4. Forest C, Czerucka D, Negrel R, Ailhaud G . Establishment of a human cell line after transformation by a plasmid containing the early region of the SV40 genome Cell Biol Int Rep 1983 7: 73–81.

    Article  CAS  Google Scholar 

  5. Tontonoz P, Singer S, Forma BM, Sarraf P, Fletcher JA, Fletcher CDM, Brun RP, Mueller E, Altiok S, Oppenheim H, Evans RM, Spiegelman BM . Terminal differentiation of human liposarcoma cells induced by ligands for peroxisome proliferator-activated receptor γ and the retinoid X receptor Proc Natl Acad Sci USA 1997 94: 237–241.

    Article  CAS  Google Scholar 

  6. Simpson JL, Landey S, New M, German J . A previously unrecognized X-linked syndrome of dysmorphia Birth Defects Orig Artic Ser 1975 11: 18–24.

    CAS  PubMed  Google Scholar 

  7. Golabi M, Rosen L . A new X-linked mental retardation-overgrowth syndrome Am J Med Genet 1984 17: 345–358.

    Article  CAS  Google Scholar 

  8. Behmel A, Pochl E, Rosenkranz W . A new X-linked dysplasia gigantism syndrome identical with Simpson dysplasia syndrome? Hum Genet 1984 67: 409–413.

    Article  CAS  Google Scholar 

  9. Pilia G, Hughes-Benzie RM, MacKenzie A, Baybayan P, Chen EY, Huber R, Neri G, Cao A, Forabosco A, Schlessinger D . Mutations in GPC3, a glypican gene, cause the Simpson-Golabi-Behmel overgrowth syndrome Nature Genet 1996 12: 241–247.

    Article  CAS  Google Scholar 

  10. Lindsay S, Ireland M, O'Brian O, Clayton-Smith J, Hurst JA, Mann J, Cole T, Sampson J, Slaney S, Schlessinger D, Burn J, Pilia G . Large scale deletions in the GPC3 gene may account for a minority of cases of Simpson–Golabi–Behmel syndrome J Med Genet 1997 34: 480–483.

    Article  CAS  Google Scholar 

  11. Hauner H, Entenmann G, Wabitsch M, Gaillard D, Ailhaud G, Negrel R, Pfeiffer EF . Promoting effect of glucocorticoids on the differentiation of human adipocyte precursor cells cultured in an chemically defined medium J Clin Invest 1989 84: 1663–1670.

    Article  CAS  Google Scholar 

  12. Wabitsch M, Braun S, Hauner H, Heinze E, Ilondo MM, Shymko R, DeMeyts P, Teller WM . Mitogenic and antiadipogenic properties of human growth hormone in human adipocyte precursor cells in primary culture Pediatr Res 1996 40: 450–456.

    Article  CAS  Google Scholar 

  13. Jensen J, Serup P, Karlsen C, Nielsen TF, Madsen OD . mRNA profiling of rat islet tumours reveals nkx 6.1 as a beta-cell-specific homeodomain transcription factor J Biol Chem 1996 271: 18749–18758.

    Article  CAS  Google Scholar 

  14. Wabitsch M, Blum WF, Muche R, Braun M, Hube F, Rascher W, Heinze E, Teller W, Hauner H . Contribution of androgens to the gender difference in leptin production in obese children and adolescents J Clin Invest 1997 100: 808–813.

    Article  CAS  Google Scholar 

  15. Chomczynski N, Sacchi N . Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chlorophorm extraction Anal Biochem 1997 162: 156–159.

    Article  Google Scholar 

  16. Pairault J, Green H . A study of the adipose conversion of suspended 3T3 cells by using glycerophosphate dehydrogenase as differentiation marker Proc Natl Acad Sci USA 1979 76: 5138–5142.

    Article  CAS  Google Scholar 

  17. Hellmér J, Arner P, Lundin A . Automatic luminometric kinetic assay of glycerol for lipolysis studies Anal Biochem 1989 177: 132–137.

    Article  Google Scholar 

  18. Blum WF, Englaro P, Hanitsch S, Juul A, Hertel NT, Müller J, Skakkebaek NE, Heiman ML, Birkett M, Attanasio AM, Kiess W, Rascher W . Plasma leptin levels in healthy children and adolescents: dependency on body mass index, body fat mass, gender, pubertal stage, and testosterone J Clin Endocrinol Metab 1997 82: 2904–2910.

    CAS  PubMed  Google Scholar 

  19. Du Manoir S, Schröck E, Bentz M, Speicher MR, Joos S, Ried T, Lichter P, Cremer T . Quantitative analysis of comparative genome hybridization Cytometry 1995 19: 27–41.

    Article  CAS  Google Scholar 

  20. Miller SA, Dykesm DD, Polesky HF . A simple salting out procedure for extracting DNA from human nucleated cells Nucleic Acids Res 1988 16: 1215.

    Article  CAS  Google Scholar 

  21. Bunge S, Fuchs S, Gal A . Simple and nonisotopic methods to detect unknown gene mutations in nucleic acids. In: Adolph KW (ed) Methods in Molecular Genetics Academic Press: Orlando, FL pp 26–39.

    Chapter  Google Scholar 

  22. Wabitsch M, Jensen PB, Blum WF, Christoffersen CT, Englaro P, Heinze E, Rascher W, Teller W, Tornqvist H, Hauner H . Insulin and cortisol promote leptin production in cultured human fat cells Diabetes 1996 45: 1435–1438.

    Article  CAS  Google Scholar 

  23. Li M, Squire JA, Weksberg R . Overgrowth syndromes and genomic imprinting: from mouse to man Clin Genet 1998 53: 165–170.

    Article  CAS  Google Scholar 

  24. Gonzalez AD, Kaya M, Shi W, Song H, Testa JR, Penn LZ, Filmus J . OCI-5/GPC3, a glypican encoded by a gene that is mutated in the Simpson–Golabi–Behmel overgrowth syndrome, induces apoptosis in a cell line-specific manner J Cell Biol 1998 141: 1407–1414.

    Article  CAS  Google Scholar 

  25. Wecksberg R, Squire JA, Templeton DM . Glypicans: a growing trend Nature Genet 1996 12: 225–227.

    Article  Google Scholar 

  26. Song HH, Shi W, Filmus J . OCI-5/rat glypican-3 binds to fibroblast growth factor-2 but not to insulin-like growth factor-2 J Biol Chem 1997 272: 7574–7577.

    Article  CAS  Google Scholar 

  27. Spiegelman BM . PPAR-γ: adipogenic regulator and thiazolidinedione receptor Diabetes 1998 47: 507–514.

    Article  CAS  Google Scholar 

  28. Tontonoz P, Hu E, Spiegelman BM . Stimulation of adipogenesis in fibroblasts by PPAR gamma 2, a lipid-activated transcription factor Cell 1994 79: 1147–1156.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Cytogenetics and CGH analysis were performed by Dr M. Djalali, Department of Medical Genetics, University of Ulm, and GPC3 mutation analysis by Ulrike Orth, Institute of Human Genetics, University of Hamburg, Germany. The authors wish to thank Simone Schmid, Uta Späth and Silvia Röderer for their excellent technical assistance. The authors gratefully acknowledge the helpful discussions and technical advices of Dr Hans Tornqvist, Department of Pediatrics, University of Lund, Sweden, and Dr Claude Marcus, Department of Pediatrics, Huddinge University Hospital, Sweden. This work was partly supported by a grant from the Deutsche Forschungsgemeinschaft (WA 1096/1-1).

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Authors and Affiliations

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

    M Wabitsch, M Braun, E Heinze & K-M Debatin

  2. Department of Orthopaedics, University of Ulm, Ulm, Germany

    RE Brenner

  3. Department of Pathology, University of Ulm, Ulm, Germany

    I Melzner & P Möller

  4. Diabetes Research Institute at the University of Duesseldorf, Duesseldorf, Germany

    H Hauner

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  1. M Wabitsch
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  2. RE Brenner
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Correspondence to M Wabitsch.

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Wabitsch, M., Brenner, R., Melzner, I. et al. Characterization of a human preadipocyte cell strain with high capacity for adipose differentiation. Int J Obes 25, 8–15 (2001). https://doi.org/10.1038/sj.ijo.0801520

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