Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptional profiling by array of human adipose-derived stem cells to study the influence of cell-secreted ECM on the osteogenic differentiation process.

ABSTRACT: The aim of this study is to characterize how the extracellular matrix secreted by adipose-derived stem cells (ADSC) during osteogenesis affects the differentiation process. Specifically, ADSC undergo osteogenesis by following similar maturational phases as bone marrow-derived stem cells. However, it is unclear how the differentiation process is the same and how it differs. We first focused on ADSC behavior by analyzing whole transcriptome changes in response to osteogenic media supplements added into the tissue culture medium. We then developed osteogenic differentiation expression profiles for the ADSC and identify key genes and pathways that serve as an osteogenesis signature. This expression signature acted as a template for comparison of how extracellular matrix (ECM) affected ADSC differentiation. We studied ADSC induced to differentiate on ECM isolated from day 16 in the differentiation process (the midpoint in osteogenesis) as well as ECM from day 11 in the differentiation process. Ultimately, we aim to dissect the relationship between cells grown on ECM as an in vitro growth substrate and cells grown on tissue culture plastic; both in the presence of osteogenic supplements. This study, will allow us to determine the extent to which ECM affects the differentiation process.

ORGANISM(S): Homo sapiens

SUBMITTER: Heather Coan

PROVIDER: E-MEXP-3340 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The methodology for the repair of critical-sized or non-union bone lesions has unpredictable efficacy due in part to our incomplete knowledge of bone repair and the biocompatibility of bone substitutes. Although human mesenchymal stem cells (hMSCs) differentiate into osteoblasts, which promote bone growth, their ability to repair bone has been unpredictable. We hypothesized that given the multi-stage process of osteogenesis, hMSC-mediated repair might be maximal at a specific time-point of healing. Utilizing a mouse model of calvarial healing, we demonstrate that the osteo-repair capacity of hMSCs can be substantially augmented by treatment with an inhibitor of peroxisome-proliferator-activated-receptor-γ, but efficacy is confined to the rapid osteogenic phase. Upon entry into the bone-remodeling phase, hMSC retention signals are lost, resulting in truncation of healing. To solve this limitation, we prepared a scaffold consisting of hMSC-derived extracellular matrix (ECM) containing the necessary biomolecules for extended site-specific hMSC retention. When inhibitor-treated hMSCs were co-administered with ECM, they remained at the injury well into the remodeling phase of healing, which resulted in reproducible and complete repair of critical-sized defects in 3 weeks. These data suggest that hMSC-derived ECM and inhibitor-treated hMSCs could be employed at optimal times to substantially and reproducibly improve bone repair. To gain insight into the superior healing potential of GW-hMSCs and also what might be accounting for their extended engraftment, microarray analyses on the RNA extracted from the calvarial tissue recovered after days 5 and 14 were performed. Equal amounts of total RNA from 4 animals per group and time point were pooled and animals receiving control (DMSO) or peroxisome proliferator-activated receptor-gamma inhibitor GW9662-treated hMSCs were compared with the assumption that murine cross-hybridization would be constant throughout the samples and thus be subtracted from the analysis.

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Transcriptional profiling by array of human adipose-derived stem cells to study the influence of cell-secreted ECM on the osteogenic differentiation process.

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