title: Decrease of apoptosis markers during adipogenic differentiation of mesenchymal stem cells from human adipose tissue.
abstract: Although the proliferation and differentiation of mesenchymal stem cells (MSCs) from adipose tissue (AT) have been widely studied, relatively little information is available on the underlying mechanism of apoptosis during the adipogenic differentiation. Thus, the aim of this study was to analyze how the expression of some apoptotic markers is affected by in vitro expansion during adipogenic differentiation of AT-MSCs. The cultures incubated or not with adipogenic medium were investigated by Western blot at 7, 14, 21, and 28 days for the production of p53, AKT, pAKT, Bax, PDCD4 and PTEN. MSCs were recognized for their immunoreactivity to MSC-specific cell types markers by immunocytochemical procedure. The effectiveness of adipogenic differentiation was assessed by staining with Sudan III and examination of adipogenic markers expression, such as PPAR-γ and FABP, at different time points by Western blot. The adipogenic differentiation medium led to the appearance, after 7 days, of larger rounded cells presenting numerous vacuoles containing lipids in which it was evident a red-orange staining, that increased in size in a time-dependent manner, parallel to an increase of the levels of expression of PPAR-γ and FABP. More than 50 % of human MSCs were fully differentiated into adipocytes within the four-week induction period. The results showed that during adipogenic differentiation of AT-MSCs the PI3K/AKT signaling pathway is activated and that p53, PTEN, PDCD4, and Bax proteins are down-regulated in time-dependent manner. Our data provide new information on the behavior of some apoptotic markers during adipogenic differentiation of AT-MSCs to apply for tissues repair and regeneration.
authors.affiliation: Department of Bio-medical Sciences, Section of Physiology, University of Catania, V.le A. Doria 6, 95125 Catania, Italy.
authors.forename: Debora
authors.initials: D
authors.lastname: Lo Furno
authors.forename: Adriana C E
authors.initials: AC
authors.lastname: Graziano
authors.forename: Silvia
authors.initials: S
authors.lastname: Caggia
authors.forename: Rosario E
authors.initials: RE
authors.lastname: Perrotta
authors.forename: Maria Stella
authors.initials: MS
authors.lastname: Tarico
authors.forename: Rosario
authors.initials: R
authors.lastname: Giuffrida
authors.forename: Venera
authors.initials: V
authors.lastname: Cardile
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000378:metabolism*; D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism*; D000328:Adult; D017209:Apoptosis / Q000235:genetics; D051017:Apoptosis Regulatory Proteins / Q000235:genetics* / Q000378:metabolism; D001391:Azo Compounds; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism; D061251:Primary Cell Culture; D015398:Signal Transduction; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D051017:Apoptosis Regulatory Proteins; D001391:Azo Compounds; D015415:Biomarkers; D047495:PPAR gamma; D025521:Tumor Suppressor Proteins; C033006:sudan III