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Genome-wide profiling of microRNAs in adipose mesenchymal stem cell differentiation and mouse models of obesity.

ABSTRACT: In recent years, there has been accumulating evidence that microRNAs are key regulator molecules of gene expression. The cellular processes that are regulated by microRNAs include e.g. cell proliferation, programmed cell death and cell differentiation. Adipocyte differentiation is a highly regulated cellular process for which several important regulating factors have been discovered, but still not all are known to fully understand the underlying mechanisms. In the present study, we analyzed the expression of 597 microRNAs during the differentiation of mouse mesenchymal stem cells into terminally differentiated adipocytes by real-time RT-PCR. In total, 66 miRNAs were differentially expressed in mesenchymal stem cell-derived adipocytes compared to the undifferentiated progenitor cells. To further study the regulation of these 66 miRNAs in white adipose tissue in vivo and their dependence on PPAR? activity, mouse models of genetically or diet induced obesity as well as a mouse line expressing a dominant negative PPAR? mutant were employed.

SUBMITTER: Bengestrate L

PROVIDER: S-EPMC3121761 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature

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Genome-wide profiling of microRNAs in adipose mesenchymal stem cell differentiation and mouse models of obesity.

Bengestrate Lena L Virtue Sam S Campbell Mark M Vidal-Puig Antonio A Hadaschik Dirk D Hahn Peter P Bielke Wolfgang W

PloS one 20110623 6

In recent years, there has been accumulating evidence that microRNAs are key regulator molecules of gene expression. The cellular processes that are regulated by microRNAs include e.g. cell proliferation, programmed cell death and cell differentiation. Adipocyte differentiation is a highly regulated cellular process for which several important regulating factors have been discovered, but still not all are known to fully understand the underlying mechanisms. In the present study, we analyzed the ...[more]

PMID: 21731698

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Project description:BackgroundRecent publications about differentiation of stem cells to germ cells have motivated researchers to make new approaches to infertility. In vitro production of germ cells improves understanding differentiation process of male and female germ cells. Due to the problem of using embryonic stem cells (ESC), it's necessary the mentioned cells be replaced with some adult multi-potent stem cells in laboratories.ObjectiveThe aim of this study was to obtain germ cells from appropriate source beyond ESC and compare differential potentials of adipocytes derived stem cells (ADMSCs) with bone marrow derived stem cells (BMMSCs).Materials and methodsTo find multi-potential entity, after providing purified ADMSCs and BMMSCs, differentiation to osteoblast and adipocyte was confirmed by using appropriate culture medium. To confirm mesenchymal lineage production superficial markers (expression of CD90 and CD44 and non-expression of CD45 and CD31) were investigated by flowcytometry. Then the cells were differentiated to germ cells in inductive medium containing retinoic acid for 7days. To evaluate germ cells characteristic markers [Dazl (Deleted in azoospermia-like), Mvh (Mouse vasa homolog gene), Stra8 (Stimulated by retinoic acid) and Scp3 (Synaptonemal complex protein 3)] flowcytometry, imunoflorescence and real time PCR were used.ResultsBoth types of cells were able to differentiate into osteoblast and adipocyte cells and presentation of stem cell superficial markers (CD90, CD44) and absence of endothelial and blood cell markers (CD31, CD45) were confirmative The flowcytometry, imunoflorescence and real time PCR results showed remarkable expression of germ cells characteristic markers (Mvh, Dazl, Stra8, and Scp3).ConclusionIt was found that although ADMSCs were attained easier and also cultured and differentiated rapidly, germ cell markers were expressed in BMMSCs significantly more than ADMSCs. This article extracted from M.Sc. thesis. (Maryam Hosseinzadeh Shirzeily).

Project description:Adult mesenchymal stem cells exert immunomodulatory effects that might improve the host response during sepsis. Knowledge on the effect of adipocyte-derived mesenchymal stem cells (ASCs) in sepsis is limited. Klebsiella (K.) pneumoniae is a common cause of gram-negative pneumonia and sepsis. The aim of this study was to determine the effect of human ASCs on the host response during pneumosepsis in mice. For this mice were infected with K. pneumoniae via the airways to induce a gradually evolving infection in the lung cumulating in pneumosepsis. One or 6 hours (h) after infection, mice were infused intravenously with ASCs or vehicle, and euthanized after 16h or 48h respectively. The effects of freshly cultured and cryopreserved ASCs were compared, the latter formulation being more clinically relevant. Intravenously administered ASCs were visualized in lung tissue by immunostaining at 1 and 3h, but not at 15h after infusion. While early after infection ASCs did not or only modestly influence bacterial loads, they reduced bacterial burdens in lungs and distant organs at 48h. ASCs reduced the lung levels of pro-inflammatory cytokines and attenuated lung pathology, but did not influence distant organ injury. ASCs strongly modified the lung transcriptome in both uninfected mice and mice with pneumosepsis, especially affecting. Cryopreserved and cultured ASCs induced largely similar effects. These data indicate that human ASCs induce profound immune modulatory effects in the lungs, resulting in reduced bacterial burdens and lung inflammation during pneumosepsis caused by a common human pathogen, suggesting that ASCs may be an adjunctive therapeutic in this condition. Experimental groups were defined as follows: (1) plac_cryo, placebo treated and K.pneumoniae infected control mice for cryopreserved MSCs; (2) plac_cult, placebo treated and K.pneumoniae infected control mice for cultured MSCs; (3) cryo, K.pneumoniae infected mice treated with cryopreserved MSCs; (4) cult, K.pneumoniae infected mice treated with cultured MSCs; (5) n_cult, non-infected control mice for cultured MSCs; (6) n_cryo, non-infected control mice for cryopreserved MSCs; (7) n, normal mouse lung baseline. In a separate experiment, mice were treated with a selective prekallikrein (PKK) directed antisense oligonucleotide (ASO) or a scrambled control ASO for 3 weeks prior to infection. Host response readouts were determined at 12 or 36 hours postinfection, including genome-wide messenger RNA profiling of lungs, or mice were followed for survival Pre-processing and quality control of the scans were performed by using the oligo method (version 1.44) and probes were annotated using the platform design info for Affymetrix Clariom_S_Mouse_HT available through Bioconductor. Array data were background corrected by Robust Multi-array Average (RMA) and quantiles-normalized. Microarray quality control was performed by means of the array quality metrics method (version 3.36.0). The occurrence of non-experimental chip effects was evaluated by the surrogate variable analysis method (version 3.28.0) and corrected using the combat method.

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Genome-wide profiling of microRNAs in adipose mesenchymal stem cell differentiation and mouse models of obesity.

Publications

Genome-wide profiling of microRNAs in adipose mesenchymal stem cell differentiation and mouse models of obesity.

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