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Development of gene expression signatures to characterize evolution of hASC cells isolated and maintained xenogeneic-free

ABSTRACT: Human adipose stem cells (hASCs) play a crucial role in the fields of regenerative medicine and tissue engineering for different reasons: the abundance of adipose tissue, their easy harvesting, the ability to multipotent differentiation and the fact that they do not trigger allogeneic blood response or secrete cytokines that act as immunosuppressants. The vast majority of protocols use animal origin reagents, with the underlying risk of transmitting infections by non-human pathogens. We have designed a protocol to isolate and maintain the properties of hASCs avoiding xenogeneic reagents. These changes not only preserve hASCs morphology, but also increase cell proliferation and maintain their stem cell marker profile. On the other hand, human serum albumin (HSA), Tryple® and human Serum (HS), do not affect hASCs multipotent differentiation ability. The amendments introduced do not trigger modifications in the transcriptional profile of hASCs, alterations in key biochemical pathways or malignization. Thus, we have proven that it is possible to isolate and maintain hASCs avoiding animal reagents and, at the same time, preserving crucial culture parameters during long term culture. Thereby we have revealed a novel and effective tool for the improvement of clinical, cell-based therapies.

ORGANISM(S): Homo sapiens

PROVIDER: GSE46314 | GEO | 2013/04/24

SECONDARY ACCESSION(S): PRJNA198722

REPOSITORIES: GEO

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Development of gene expression signatures to characterize evolution of hASC cells isolated and maintained xenogeneic-free

Project description:Human adipose stem cells (hASCs) play a crucial role in the fields of regenerative medicine and tissue engineering for different reasons: the abundance of adipose tissue, their easy harvesting, the ability to multipotent differentiation and the fact that they do not trigger allogeneic blood response or secrete cytokines that act as immunosuppressants. The vast majority of protocols use animal origin reagents, with the underlying risk of transmitting infections by non-human pathogens. We have designed a protocol to isolate and maintain the properties of hASCs avoiding xenogeneic reagents. These changes not only preserve hASCs morphology, but also increase cell proliferation and maintain their stem cell marker profile. On the other hand, human serum albumin (HSA), TrypleM-BM-. and human Serum (HS), do not affect hASCs multipotent differentiation ability. The amendments introduced do not trigger modifications in the transcriptional profile of hASCs, alterations in key biochemical pathways or malignization. Thus, we have proven that it is possible to isolate and maintain hASCs avoiding animal reagents and, at the same time, preserving crucial culture parameters during long term culture. Thereby we have revealed a novel and effective tool for the improvement of clinical, cell-based therapies. Gene expression in human adipose stem cells isolated and maintained with human reagents were measured immediately after their isolation, and at passages 1, 3 and 5. Pools from the same 8 healthy donors were used to measure the 4 points of growth.

2013-04-24 | E-GEOD-46314 | biostudies-arrayexpress

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2010-09-30 | GSE20697 | GEO

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Project description:Human abdominal adipose tissue was obtained with informed consent from a 33-year old Caucasian female (BMI = 32.96 Kg/m2) undergoing lipoaspiration. Adipose stromal cells (hASCs) were isolated and differentiated into adipocytes in vitro. Two technical replicates from 9 time points relative to induction of adipogenesis (day 0). Also, one sample from pre-adipocytes (day -2) grown without FGF.

2010-09-30 | E-GEOD-20699 | biostudies-arrayexpress

Expression profiling of human adipose stromal cell (hASC) adipogenesis

Project description:Human abdominal adipose tissue was obtained with informed consent from a 33-year old Caucasian female (BMI = 32.96 Kg/m2) undergoing lipoaspiration. Adipose stromal cells (hASCs) were isolated and differentiated into adipocytes in vitro. Two technical replicates from 9 time points realtive to induction of adipogenesis (day 0). Also, two technical replicated from pre-adipocytes (day -2) grown without FGF. Two individual replicates (day 1 and day 7) were removed due to failed hybridizations.

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Human dental pulp cells cultured in FBS vs. chemically defined conditions

Project description:Human dental pulp cells (hDPCs) are one of the promising resources for regenerative medicine and tissue engineering, including derivation of induced pluripotent stem cells (iPSCs). However, our current protocol uses reagents of animal origin, mainly fetal bovine serum (FBS) with potential risk of infectious diseases and unwanted immunogenicity. This time, we designed a chemically defined protocol to isolate and maintain the growth and differentiation potentials of hDPCs.

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TGFβ1 determines the fate of human adipose-derived stromal cells by restraining their immaturity and white/beige adipocyte potentials

Project description:Human adipose-derived stem/stromal cells (hASCs) can differentiate into a large broad of specialized cell types to ensure homeostasis of tissues. Thus, they are drawing increasing attention in cell therapy and regenerative medicine. However, culture conditions are still critical to ensure their regenerative capabilities. Here we compared Standard conditions (αMEM containing 10% fetal bovine serum) and Endothelial cell Growth Medium 2 (EGM2) containing few serum (2% v/v) for expansion of hASCs. Both types of hASCs were exhaustively characterized by high throughput studies, Gene Set Enrichment Analyses (GSEA) and differentiation potentials experiments. EGM2-hASCs showed enhanced multipotency and their phenotype was more immature than Standard-hASCs. The adipogenic potential of EGM2-hASCs was clearly more extented, including toward the thermogenic beige adipocyte fate. Data from microarray and GSEA highlighted Transforming Growth Factor β1 (TGFβ1) as upstream factor influencing the becoming of Standard-hASCs which is consistent with higher TGFβ1 content in Standard medium. This was associated with nuclear SMAD3 localization and higher expression of its active form in Standard-hASCs. Moreover, these cells were primed into osteoblast, chondroblast and Vascular Smooth Muscle (VSM) lineages at the expense of their adipogenic potential. Their treatment with TGFβ1 receptor inhibitors resulted in a cytoplasmic re-localization of SMAD3 and a decrease of VSM and osteoblastic lineages markers, increasing in turn their beige adipogenic potential. Therefore, TGFβ1 is a key factor committing hASCs toward osteo-chondroblastic and VSM lineages at the expanse of their beige adipogenic potential. Thanks to low TGFβ1 content, EGM2 medium improves the maintenance of uncommitted hASCs with strong beige adipocyte potential. We used microarrays to unravel differential gene expressions between human ASCs cultured in standard conditions or in EGM2 medium.

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Project description:Human adipose-derived stem cells (hASCs) are a highly attractive source in bone tissue engineering.We found that MIR31HG significantly promote the osteogenic differentiation of hASCs. However, the underlying mechanisms still await investigations. Thus the alteration of gene expression induced by MIR31HG knockdown was investigated. We used microarrays to detail the global programme of gene expression when MIR31HG was knocked down in hASCs and identified distinct classes of dn-regulated genes during this process.

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Genome-wide gene expression during osteogenic and myogenic differentiation from adipose- derived stem cells.

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2013-10-03 | E-GEOD-37329 | biostudies-arrayexpress

Genome-wide gene expression during osteogenic and myogenic differentiation from adipose- derived stem cells.

2013-10-03 | GSE37329 | GEO

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