Gene expression profile of osteogenically differentiated human adipose-derived stem cells
ABSTRACT: Adipose tissue harbours a significant number of multipotent adult stem cells of mesenchymal origin known as adipose-derived stem cells (ADSCs). Broad differentiation potential and convenient accessibility of ADSCs make them an attractive source of adult mesenchymal stem cell for regenerative medicine and cell developmental plasticity research. Genome-wide microarray expression profiling was performed to identify genes deregulated during osteogenic differentiation of ADSCs to evaluate developmental plasticity of these cells. Dynamics of epigenetic modifications were analyzed in parallel and associated with the gene expression profile. Gene expression profile was analyzed in adipose-derived stem cells (ADSCs) differentiated into osteogenic lineage from 3 donors and compared to undifferentiated cells from the same donors.
Project description:Mesenchymal stem cells (MSCs) are the multipotent stem cells of adult human tissues that have the ability to replicate with high proliferative rates and are responsible for the tissue renewal and regeneration. Effective osteogenic differentiation of adipose-derived stem cells (ADSCs) encourages clinical application of the cells in bone regeneration. Regeneration of damaged joints is highly dependent on the presence of MSC in synovia. Synovial membrane-derived MSCs (SM-MSCs) show good multi-lineage differentiation potential, low degree of invasiveness, and are considered as an alternative treatment strategy for arthritis-damaged tissues. Studies in MSCs from different sources identified several miRNAs important for osteogenesis. Less is known about the participation of particular miRNAs in osteogenic differentiation of SM-MSCs, the potential source of MSCs for authologous cell therapy in arthritis. In the present study, the changes in miRNA expression profile occurring during osteogenic differentiation were analyzed in human ADSCs and SM-MSCs by microarray-based and quantitative PCR approaches. We aimed at the identification of miRNAs involved in the maintenance of these MSCs and regulation of osteogenic differentiation of ADSCs and SM-MSCs. Overall design: MiRNA expression profile was analyzed in synovial membrane-derived (SM-MSCs, 3 cell lines) and adipose-derived (ADSCs, 4 cell lines) stem cells differentiated into osteogenic lineage and compared to undifferentiated cells from the same donors. Two ADSCs cell lines of adipogenic differentiation were included for comparison.
Project description:we analyzed the expression level change of transcription factors in adipose derived stem cells during osteogenic differentiation and found a candidate target gene, Sox11. We defined that Sox11 suppresses osteogenic differentiation through overexpression and knock down of Sox11. total RNA obtained from adipose derived stem cells subjected to 1,3,6,10 or 14 days in osteogenic differentiation compared to undifferentiated control adipose derived stem cells.
Project description:To see the effect of zoledronate on the osteogenic differentiation of human mesenchymal stem cells. Cells were first cultured in normal media until enough cells were obtained. Then the cells were differentiated into ostepbalst under the effect of low concentration of zoledronate.
Project description:Background: Tissue regeneration and recovery in the adult body depends on self-renewal and differentiation of stem and progenitor cells. Mesenchymal stem cells (MSCs) that have the ability to differentiate into various cell types, have been isolated from the stromal fraction of virtually all tissues. However, little is known about the true identity of MSCs. MSC populations exhibit great tissue-, location- and patient-specific variation in gene expression and are heterogeneous in cell composition. Methodology/Principal findings: Our aim was to analyze the dynamics of differentiation of two closely related stromal cell types, adipose tissue-derived MSCs and dermal fibroblasts (FBs) along adipogenic, osteogenic and chondrogenic lineages using multiplex RNA-seq technology. We found that undifferentiated donor-matched MSCs and FBs are distinct populations that stay different upon differentiation into adipocytes, osteoblasts and chondrocytes. The changes in lineage-specific gene expression occur early in differentiation and persist over time in both MSCs and FBs. Further, MSCs and FBs exhibit similar dynamics of adipogenic and osteogenic differentiation but different dynamics of chondrogenic differentiation. Conclusion: Our findings suggest that stromal stem cells including adipose-derived MSCs and dermal FBs exploit different molecular mechanisms of differentiation to reach a common cell fate. The early mechanisms of differentiation are lineage-specific and are similar for adipogenic and osteogenic differentiation but are distinct for chondrogenic differentiation between MSCs and FBs. A total of 91 samples were analyzed by multiplex RNA-seq. Samples represented replicates from two patients, two cell types and three differentiation protocols, as indicated by the sample annotation. 5 barcodes were unused, but the corresponding FASTQ files are included for completeness.
Project description:The time course experiment was designed as follow: (1) Mesenchymal stem cell (indifferentiated cells)/ time 0h; (2) After 24 h of osteogenic induction; (3) After 48 h of osteogenic induction (4) After 7 days of osteogenic induction.
Project description:Adipose-derived and bone-marrow-derived mesenchymal stem cells were collected from 3 pigs and cultivated in vitro up to 3 passages. At passage 3 cells were cultured to 80% confluence and induced to differentiate in adipose and bone. Cell were harvested at 0 day of differentiation (dd) or pre-differentiation, at 2, 7, and 21dd for RNA extraction. The RNA was used for a large microarray analysis using a specific pig oligo-array with >10,000 annotated genes. The main aim of the microarray analysis was to directly compare the two transcriptomics adaptation of the two mesenchymal stem cells during osteogenic and adipogenic differentiation The mesenchymal stem cells were harvested at 0, 2, 7, and 21 day of differentiation (dd). A dye-swap reference design (reference = mixture of RNA from several porcine tissues) was used.
Project description:The in vitro osteogenic differentiation of hAEC, hAFSC and hBMSC have been reported.The final tissue-forming potential of all three cell types may vary in terms of different anatomical origin and molecular response to osteogenic induction. We used microarrays to detail the global genes expression profiles of hAEC, hAFSC and hBMSC before and after osteogenic induction in vitro. Global gene expression profiles of hAECs, hBMSCs and hAFMSCs were evaluated before and after 7-day osteogenic induction in vitro. Samples were subjected to gene expression analysis using the Affymetrix human HTA2.0 microarray.
Project description:Dedifferentiated fat (DFAT) cells, established in vitro from mature adipocytes, exhibit certain properties of multipotent mesenchymal stem/stromal cells (MSCs), such as the ability to differentiate into multiple mesenchymal lineages. Although DFAT cells exhibit properties of proliferative progeny, at present there is only limited knowledge about their MSC-specific characteristics because those cells are considered to be potential artifacts of cell culture. To elucidate the identity of DFAT cells, we compared gene expression profiles of human DFAT cells and adipose-derived stem/stromal cells (ADSCs) established using adipose tissue from the same donors. Microarray analysis showed that global mRNA expression profiles of human DFAT cells were very similar to those of ADSCs, a representative MSC, despite being committed adipocyte progenitors. Overall design: Subcutaneous adipose tissues that were obtained during surgical operation for non-malignant disease were donated by 3 patients after obtaining informed consent. Three sets of DFAT cells and ADSCs, each derived from adipose tissue from the same donor were used for RNA extraction and subsequent microarray analysis.
Project description:To identify the key microRNAs (miRNAs) of hMSCs required for fate determination, miRNA profiling was performed with hMSCs from three different sources including adipose-derived stem cells (ADSCs), bone-marrow-derived stem cells (BMSCs), and umbilical cord-derived stem cells (UCSCs) versus fibroblasts, a more differentiated mesenchymal cell types. We compared the expression profiles of two different donors per hMSCs to that of fibroblasts. All hMSCs were used for profiling at passage 3-6.
Project description:Extracellular nucleotides are potent signaling molecules mediating cell-specific biological functions. We previously demonstrated that adenosine 5'-triphosphate (ATP) inhibits the proliferation while stimulating the migration, in vitro and in vivo, of human bone marrow-derived mesenchymal stem cells (BM-hMSC). Here, we investigated the effects of ATP on BM-hMSC differentiation capacity. Molecular analysis showed that ATP treatment modulated the expression of several genes (e.g. wnt-pathway-related genes) governing osteoblastic and adipogenic differentiation of MSCs. Functional studies demonstrated that ATP, under specific culture conditions, stimulated adipogenic and osteogenic differentiation by significantly increasing the lipid accumulation and the expression levels of the adipogenic master gene PPARγ (peroxisome proliferator activated receptor-gamma) and by promoting the mineralization and the expression of the osteoblast-related gene RUNX2 (Runt-related transcription factor 2), respectively. BM-hMSCs cells were transiently exposed to ATP 1mM for 24 hours (ATP pre-treatment) before starting differentiation induction. Then, BM-hMSCs were cultured under adipogenic/osteogenic conditions. Gene Expression Profile was performed on differentiate cells after 3 weeks of induction culture.