Thiazolidinediones and glucocorticoids synergistically induce differentiation of human adipose tissue stromal cells: Biochemical, cellular, and molecular analysis

While adipocyte differentiation has been studied extensively in murine cultures, the lack of a readily available preadipocyte model has hindered equivalent studies in man. We describe methods for the isolation and culture of primary human stromal cells from surgical adipose tissue specimens. In vitr...

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Bibliographic Details
Published in:	Metabolism, clinical and experimental Vol. 50; no. 4; pp. 407 - 413
Main Authors:	Halvorsen, Yuan-Di C., Bond, Arden, Sen, Anindita, Franklin, Dawn M., Lea-Currie, Y.Renee, Sujkowski, Danuta, Ellis, P.Nicole, Wilkison, William O., Gimble, Jeffrey M.
Format:	Journal Article
Language:	English
Published:	New York, NY Elsevier Inc 01-04-2001 Elsevier
Subjects:	Adipose Tissue - cytology Adipose Tissue - drug effects Adipose Tissue - metabolism Biological and medical sciences Cell Differentiation - drug effects Cell differentiation, maturation, development, hematopoiesis Cell physiology Coloring Agents Fundamental and applied biological sciences. Psychology Glucocorticoids - pharmacology Glycerolphosphate Dehydrogenase - metabolism Humans Immunoblotting In Vitro Techniques Leptin - biosynthesis Lipectomy Lipolysis - drug effects Molecular and cellular biology Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger - biosynthesis Stromal Cells - drug effects Stromal Cells - metabolism Thiazoles - pharmacology Human Adipocyte Cell culture Adipose tissue Steroid hormone Biochemistry Cell differentiation Glucocorticoid In vitro Biological activity Adrenal hormone Stromal cell Molecular biology
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Summary:	While adipocyte differentiation has been studied extensively in murine cultures, the lack of a readily available preadipocyte model has hindered equivalent studies in man. We describe methods for the isolation and culture of primary human stromal cells from surgical adipose tissue specimens. In vitro, the stromal cells rapidly differentiate in response to a combination of adipogenic agents. Among these, glucocorticoids and thiazolidinediones act together to induce the formation of lipid vacuoles within the cells. These morphologic changes accompany the increased expression of 2 characteristic adipocyte proteins, the cytoplasmic enzyme glycerol phosphate dehydrogenase (GPDH) and the secreted cytokine leptin. Likewise, stromal cell differentiation results in elevated mRNA levels for the fatty acid binding protein aP2 and the adipogenic regulatory transcription factors CCAAT/enhancer binding protein α (C/EBPα) and peroxisome proliferator-activated receptor γ (PPARγ) in addition to leptin. The in vitro differentiated stromal cells exhibit a lipolytic response to β-adrenergic agonists, comparable to that reported with primary human adipocytes. These studies demonstrate the validity of human adipose tissue-derived stromal cells as a reliable in vitro model for investigations of adipocyte metabolism in humans.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0026-0495 1532-8600
DOI:	10.1053/meta.2001.21690