A novel technique to propagate primary human preadipocytes without loss of differentiation capacity

Obesity
T SkurkH Hauner

Abstract

The study of human preadipocytes is hampered by the limited availability of adipose tissue and low yield of cell preparation. Proliferation of preadipocytes using common protocols, including fetal bovine serum (FBS), results in a markedly reduced differentiation capacity. Therefore, we were interested in developing an improved culture system that allows the proliferation of human preadipocytes without loss of differentiation capacity. Adipose tissue samples were taken from subjects undergoing elective abdominal surgery. Cells were seeded at various densities and cultured using different formulations of proliferation media including factors such as fibroblast growth factor-2 (basic fibroblast growth factor), epidermal growth factor, insulin, and FBS either alone or in combination. Cells were counted and induced to differentiate after confluence. After complete differentiation, cells were harvested, and glycerol-3-phosphate dehydrogenase activity was measured. Cells were subcultured for up to five passages. The proliferation medium with 4 growth factors (PM4), consisting of 2.5% FBS, 10 ng/mL epidermal growth factor, 1 ng/mL basic fibroblast growth factor, and 8.7 muM insulin, resulted in lower doubling times at all seeding densi...Continue Reading

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Citations

May 10, 2013·The Journals of Gerontology. Series A, Biological Sciences and Medical Sciences·Maria C MitterbergerWerner Zwerschke
Feb 14, 2016·Biochemical and Biophysical Research Communications·Diana RöhrbornJürgen Eckel
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May 12, 2021·Stem Cell Research & Therapy·Christian PlonerGerhard Pierer
Jul 13, 2021·American Journal of Physiology. Endocrinology and Metabolism·Julie VionDominique Langin
Apr 2, 2009·Current Opinion in Organ Transplantation

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