Expression analysis of human adipose-derived stem cells during in vitro differentiation to an adipocyte lineage

Satish, L and Krill-Burger, JM and Gallo, PH and Etages, SD and Liu, F and Philips, BJ and Ravuri, S and Marra, KG and LaFramboise, WA and Kathju, S and Rubin, JP (2015) Expression analysis of human adipose-derived stem cells during in vitro differentiation to an adipocyte lineage. BMC Medical Genomics, 8 (1).

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Abstract

Background: Adipose tissue-derived stromal stem cells (ASCs) represent a promising regenerative resource for soft tissue reconstruction. Although autologous grafting of whole fat has long been practiced, a major clinical limitation of this technique is inconsistent long-term graft retention. To understand the changes in cell function during the transition of ASCs into fully mature fat cells, we compared the transcriptome profiles of cultured undifferentiated human primary ASCs under conditions leading to acquisition of a mature adipocyte phenotype. Methods: Microarray analysis was performed on total RNA extracted from separate ACS isolates of six human adult females before and after 7 days (7 days: early stage) and 21 days (21 days: late stage) of adipocyte differentiation in vitro. Differential gene expression profiles were determined using Partek Genomics Suite Version 6.4 for analysis of variance (ANOVA) based on time in culture. We also performed unsupervised hierarchical clustering to test for gene expression patterns among the three cell populations. Ingenuity Pathway Analysis was used to determine biologically significant networks and canonical pathways relevant to adipogenesis. Results: Cells at each stage showed remarkable intra-group consistency of expression profiles while abundant differences were detected across stages and groups. More than 14,000 transcripts were significantly altered during differentiation while ~6000 transcripts were affected between 7 days and 21 days cultures. Setting a cutoff of +/-two-fold change, 1350 transcripts were elevated while 2929 genes were significantly decreased by 7 days. Comparison of early and late stage cultures revealed increased expression of 1107 transcripts while 606 genes showed significantly reduced expression. In addition to confirming differential expression of known markers of adipogenesis (e.g., FABP4, ADIPOQ, PLIN4), multiple genes and signaling pathways not previously known to be involved in regulating adipogenesis were identified (e.g. POSTN, PPP1R1A, FGF11) as potential novel mediators of adipogenesis. Quantitative RT-PCR validated the microarray results. Conclusions: ASC maturation into an adipocyte phenotype proceeds from a gene expression program that involves thousands of genes. This is the first study to compare mRNA expression profiles during early and late stage adipogenesis using cultured human primary ASCs from multiple patients.

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Details

Item Type:	Article
Status:	Published
Creators/Authors:	Creators Email Pitt Username ORCID Satish, L las238@pitt.edu LAS238 Krill-Burger, JM Gallo, PH phg7@pitt.edu PHG7 Etages, SD Liu, F fal10@pitt.edu FAL10 Philips, BJ bjp24@pitt.edu BJP24 Ravuri, S Marra, KG kgm5@pitt.edu KGM5 0000-0002-8437-4864 LaFramboise, WA wal9@pitt.edu WAL9 0000-0002-6024-810X Kathju, S Rubin, JP jpr5@pitt.edu JPR5
Centers:	Other Centers, Institutes, Offices, or Units > McGowan Institute for Regenerative Medicine
Date:	24 July 2015
Date Type:	Publication
Journal or Publication Title:	BMC Medical Genomics
Volume:	8
Number:	1
DOI or Unique Handle:	10.1186/s12920-015-0119-8
Schools and Programs:	School of Medicine > Pathology School of Medicine > Surgery
Refereed:	Yes
Date Deposited:	17 Aug 2016 13:46
Last Modified:	11 Sep 2023 14:55
URI:	http://d-scholarship.pitt.edu/id/eprint/29227

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