Plastic Surgery Research Council
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Presenter: Peter J. Rubin, MD
Co-Authors: Krill-Burger M; Philips B; Gallo P; Des Estages S; Liu F; Ravuri S; Laframboise WA; Marra KG; Kathju S; Rubin JP
university of pittsburgh

Introduction: Adipose tissue-derived mesenchymal stromal cells (ASCs) represent a promising regenerative source for soft tissue reconstruction. One limitation of this technique is the poor long-term retention in clinical practice. The variable success rates may stem from different functional properties of heterogeneous cell populations found within the stromal vascular fraction.

Materials and Methods: To better understand signaling pathway differences and implications for changes in cell function during the transition of ASCs into fully mature and functional adipocytes/fat, we compared the mRNA profiles of undifferentiated (0d) and differentiated (7d and 21d) primary cultured human ASCs under conditions of adipocyte lineage. Molecular profiling was performed on total RNA extracted from 0d, 7d and 21d cell populations obtained from six human adult female patients using Affymetrix Human U133 plus 2.0 gene arrays.

Results: Data were analyzed using Partek software, and two-way ANOVA was performed to determine statistically significant differences based on cell grouping. followed by post-hoc testing correcting for false discovery rates (<5%). Each grouping of cells was substantially different from each other and undifferentiated cells were markedly different from the cells cultured for either 7d or 21d by 14,000 transcripts. Between 7d and 21d cultures there were 6,000 different transcripts. Comparing 7d vs undifferentiated cells, 1350 genes were up-regulated (2fold and above) and 2929 genes were down-regulated (0.5fold and below). Comparing 21d vs 7d, 1107 genes were up-regulated and 606 genes were down-regulated. In addition to genes known to be key markers of adipogenesis (e.g., FABP4, PPAR?), several genes and associated signaling pathways not known to be involved in regulating adipogenesis were identified, and may represent novel adipogenic mediators. QRT-PCR performed on select genes validated the microarray results.

Discussion: To our knowledge, this is the first study to assess and compare early and late mRNA expression profiles during adipogenesis using primary cultured human ASC s from multiple patients.

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