Identity, proliferation capacity, genomic stability and novel senescence markers of mesenchymal stem cells isolated from low volume of human bone marrow
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE68933 | GEO | 2016/02/29
SECONDARY ACCESSION(S): PRJNA284132
REPOSITORIES: GEO
Similar Datasets
Project description:Human bone marrow mesenchymal stem cells (hBM-MSCs) hold promise for treating incurable diseases and repairing of damaged tissues. However, hBM-MSCs face the disadvantages of painful invasive isolation and limited cell numbers. In this study we assessed characteristics of MSCs isolated from residual human bone marrow transplantation material and expanded to clinically relevant numbers at passages 3-4 and 6-7. Results indicated that early passage hBM-MSCs are genomically stable and retain identity and high proliferation capacity. Despite the chromosomal stability, the cells became senescent at late passages, paralleling the slower proliferation, altered morphology and immunophenotype. By qRT-PCR array profiling, we revealed 13 genes and 33 miRNAs significantly differentially expressed in late passage cells, among which 8 genes and 30 miRNAs emerged as potential novel biomarkers of hBM-MSC aging. Functional analysis of genes with altered expression showed strong association with biological processes causing cellular senescence. Altogether, this study revives hBM as convenient source for cellular therapy. Potential novel markers provide new details for better understanding the hBM-MSC senescence mechanisms, contributing to basic science, facilitating the development of cellular therapy quality control, and providing new clues for human disease processes since senescence phenotype of the hematological patient hBM-MSCs only very recently has been revealed.
| S-EPMC4905439 | biostudies-literature
Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series
2016-02-29 | E-GEOD-68933 | biostudies-arrayexpress
Project description:Human bone marrow mesenchymal stem cells (hBM-MSCs) hold promise for treating diseases for which currently no therapeutic options exist. High quantities of MSCs are needed, requiring extensive cell expansion in long-term culture. However, the fundamental biological properties of MSCs can be altered by culture conditions. In this study, hBM-MSCs were isolated from residual human bone marrow (hBM) material and expanded to clinically relevant numbers at passage 3-4. The cells had normal morphology, proliferation rate, immunophenotype and karyotype. Despite the chromosomal stability, after additional three passages (P6-P7) hBM-MSCs entered senescence state, confirmed by SA-β-galactosidase staining and paralleling the slower proliferation, altered morphology and imunophenotype. qRT-PCR array profiling revealed 3 out of 86 genes significantly (p <0.05) differentially (â¥2 fold) expressed in the late passage cells compared to the early passage cells. qPCR gene expression profiling. hBM-MSCs from 3 (1 male and 2 females) adult donors (age 30±7.21 years) were used. Each sample was tested in technical duplicate. 6 samples of early passage (P3-P4) hBM-MSCs were grouped to CONTROL group and 6 samples of late passage (P6-P7) hBM-MSCs were grouped to TEST group. The data were analyzed using web-based RT2 Profiler PCR Array Data Analysis v3.5 software (Qiagen)
2016-02-29 | E-GEOD-68926 | biostudies-arrayexpress
Project description:Human bone marrow mesenchymal stem cells (hBM-MSCs) hold promise for treating diseases for which currently no therapeutic options exist. High quantities of MSCs are needed, requiring extensive cell expansion in long-term culture. However, the fundamental biological properties of MSCs can be altered by culture conditions. In this study, hBM-MSCs were isolated from residual human bone marrow (hBM) material and expanded to clinically relevant numbers at passage 3-4. The cells had normal morphology, proliferation rate, immunophenotype and karyotype. Despite the chromosomal stability, after additional three passages (P6-P7) hBM-MSCs entered senescence state, confirmed by SA-β-galactosidase staining and paralleling the slower proliferation, altered morphology and imunophenotype. qRT-PCR array profiling revealed 10 out of 86 genes significantly (p <0.05) differentially (â¥2 fold) expressed in the late passage cells compared to the early passage cells qPCR gene expression profiling. hBM-MSCs from 3 (1 male and 2 females) adult donors (age 30±7.21 years) were used. Each sample was tested in technical duplicate. 6 samples of early passage (P3-P4) hBM-MSCs were grouped to CONTROL group and 6 samples of late passage (P6-P7) hBM-MSCs were grouped to TEST group. The data were analyzed using web-based RT2 Profiler PCR Array Data Analysis v3.5 software (Qiagen)
2016-02-29 | E-GEOD-68931 | biostudies-arrayexpress
Project description:Human bone marrow mesenchymal stem cells (hBM-MSCs) hold promise for treating diseases for which currently no therapeutic options exist. High quantities of MSCs are needed, requiring extensive cell expansion in long-term culture. However, the fundamental biological properties of MSCs can be altered by culture conditions. In this study, hBM-MSCs were isolated from residual human bone marrow (hBM) material and expanded to clinically relevant numbers at passage 3-4. The cells had normal morphology, proliferation rate, immunophenotype and karyotype. Despite the chromosomal stability, after additional three passages (P6-P7) hBM-MSCs entered senescence state, confirmed by SA-β-galactosidase staining and paralleling the slower proliferation, altered morphology and imunophenotype. qRT-PCR array profiling revealed 33 out of 420 miRNAs significantly (p <0.05) differentially (â¥2 fold) expressed in the late passage cells compared to the early passage cells. qPCR miRNA expression profiling. hBM-MSCs from 3 (1 male and 2 females) adult donors (age 30±7.21 years) were used. Each sample was tested in technical duplicate. 6 samples of early passage (P3-P4) hBM-MSCs were grouped to CONTROL group and 6 samples of late passage (P6-P7) hBM-MSCs were grouped to TEST group. The data were analysed using web-based miScript miRNA PCR Array Data Analysis software (Qiagen)
2016-02-29 | E-GEOD-68932 | biostudies-arrayexpress
Project description:Human bone marrow mesenchymal stem cells (hBM-MSCs) hold promise for treating diseases for which currently no therapeutic options exist. High quantities of MSCs are needed, requiring extensive cell expansion in long-term culture. However, the fundamental biological properties of MSCs can be altered by culture conditions. In this study, hBM-MSCs were isolated from residual human bone marrow (hBM) material and expanded to clinically relevant numbers at passage 3-4. The cells had normal morphology, proliferation rate, immunophenotype and karyotype. Despite the chromosomal stability, after additional three passages (P6-P7) hBM-MSCs entered senescence state, confirmed by SA-β-galactosidase staining and paralleling the slower proliferation, altered morphology and imunophenotype. qRT-PCR array profiling revealed 33 out of 420 miRNAs significantly (p <0.05) differentially (≥2 fold) expressed in the late passage cells compared to the early passage cells.
2016-02-29 | GSE68932 | GEO
Project description:Human bone marrow mesenchymal stem cells (hBM-MSCs) hold promise for treating diseases for which currently no therapeutic options exist. High quantities of MSCs are needed, requiring extensive cell expansion in long-term culture. However, the fundamental biological properties of MSCs can be altered by culture conditions. In this study, hBM-MSCs were isolated from residual human bone marrow (hBM) material and expanded to clinically relevant numbers at passage 3-4. The cells had normal morphology, proliferation rate, immunophenotype and karyotype. Despite the chromosomal stability, after additional three passages (P6-P7) hBM-MSCs entered senescence state, confirmed by SA-β-galactosidase staining and paralleling the slower proliferation, altered morphology and imunophenotype. qRT-PCR array profiling revealed 10 out of 86 genes significantly (p <0.05) differentially (≥2 fold) expressed in the late passage cells compared to the early passage cells
2016-02-29 | GSE68931 | GEO
Project description:Human bone marrow mesenchymal stem cells (hBM-MSCs) hold promise for treating diseases for which currently no therapeutic options exist. High quantities of MSCs are needed, requiring extensive cell expansion in long-term culture. However, the fundamental biological properties of MSCs can be altered by culture conditions. In this study, hBM-MSCs were isolated from residual human bone marrow (hBM) material and expanded to clinically relevant numbers at passage 3-4. The cells had normal morphology, proliferation rate, immunophenotype and karyotype. Despite the chromosomal stability, after additional three passages (P6-P7) hBM-MSCs entered senescence state, confirmed by SA-β-galactosidase staining and paralleling the slower proliferation, altered morphology and imunophenotype. qRT-PCR array profiling revealed 3 out of 86 genes significantly (p <0.05) differentially (≥2 fold) expressed in the late passage cells compared to the early passage cells.
2016-02-29 | GSE68926 | GEO
Project description:TSC1 is a tumor suppressor that inhibits cell growth via negative regulation of the mammalian target of rapamycin complex (mTORC1). TSC1 mutations are associated with Tuberous Sclerosis Complex (TSC), characterized by multiple benign tumors of mesenchymal and epithelial origin. TSC1 modulates self-renewal and differentiation in hematopoietic stem cells; however, its effects on mesenchymal stem cells (MSCs) are unknown. We investigated the impact of Tsc1 inactivation in murine bone marrow (BM)-MSCs, using tissue-specific, transgelin (Tagln)-mediated cre-recombination, targeting both BM-MSCs and smooth muscle cells. Tsc1 mutants were viable, but homozygous inactivation led to a dwarfed appearance with TSC-like pathologies in multiple organs and reduced survival. In young (28 day old) mice, Tsc1 deficiency-induced significant cell expansion of non-hematopoietic BM in vivo, and MSC colony-forming potential in vitro, that was normalized upon treatment with the mTOR inhibitor, everolimus. The hyperproliferative BM-MSC phenotype was lost in aged (1.5 yr) mice, and Tsc1 inactivation was also accompanied by elevated ROS and increased senescence. ShRNA-mediated knockdown of Tsc1 in BM-MSCs replicated the hyperproliferative BM-MSC phenotype and led to impaired adipogenic and myogenic differentiation. Our data show that Tsc1 is a negative regulator of BM-MSC proliferation and support a pivotal role for the Tsc1-mTOR axis in the maintenance of the mesenchymal progenitor pool.
| S-EPMC7565438 | biostudies-literature
Project description:Cell identity genes are distinct from other genes with respect to the epigenetic mechanisms to activate their transcription, e.g. by super-enhancers and broad H3K4me3 domains. However, it remains unclear whether their post-transcriptional regulation is also unique. We performed a systematic analysis of transcriptome-wide RNA stability in nine cell types and found that unstable transcripts were enriched in cell identity-related pathways while stable transcripts were enriched in housekeeping pathways. Joint analyses of RNA stability and chromatin state revealed significant enrichment of super-enhancers and broad H3K4me3 domains at the gene loci of unstable transcripts. Intriguingly, the RNA m6A methyltransferase, METTL3, preferentially binds to chromatin at super-enhancers, broad H3K4me3 domains and their associated genes. METTL3 binding intensity is positively correlated with RNA m6A methylation and negatively correlated with RNA stability of cell identity genes, probably due to co-transcriptional m6A modifications promoting RNA decay. Nanopore direct RNA-sequencing showed that METTL3 knockdown has a stronger effect on RNA m6A and mRNA stability for cell identity genes. Our data suggest a run-and-brake model, where cell identity genes undergo both frequent transcription and fast RNA decay to achieve precise regulation of RNA expression.
| S-EPMC10325912 | biostudies-literature