Generation and Characterization of an immortalized human mesenchymal stromal cell line model
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PROVIDER: PRJNA207288 | ENA |
REPOSITORIES: ENA
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Project description:Human mesenchymal stromal cells (hMSCs) show great potential for clinical and experimental use. Self-renewal and multipotent differentiation capacity are key elements in the increasing interest of this cell type. However, a disadvantage of primary hMSCs is the limited in vitro lifespan, but this can be overcome by introduction of the catalytic subunit of human telomerase reverse transcriptase (TERT). In this paper, the generation and characterization of a TERT-immortalized, non-tumorigenic human bone marrow-derived stromal mesenchymal model cell line is described. The resulting cell line, iMSC#3, maintained capacity to differentiate into osteoblasts and adipocytes, and growth characteristics comparable to primary hMSCs into after long-term culturing (155 population doublings). A detailed characterization of the mRNA- and microRNA transcriptomes during adipocyte differentiation also showed that the iMSC#3 recapitulates this process at the molecular level. Furthermore, iMSC#3 showed no major differences from hMSCs regarding surface marker expression. The cell line had a normal karyotype, and high-resolution array comparative genomic hybridization confirmed that the cells in general had normal copy number. The gene expression profiles of immortalized and primary hMSCs were also similar, whereas the corresponding methylation profiles were more diverse. In conclusion, a human bone marrow-derived stromal cell line was developed, which can be used for basic studies of mesenchymal cells and to functionally study transgenes that might play a role in oncogenic transformation.
2014-12-31 | GSE47646 | GEO
Project description:Human mesenchymal stromal cells (hMSCs) show great potential for clinical and experimental use. Self-renewal and multipotent differentiation capacity are key elements in the increasing interest of this cell type. However, a disadvantage of primary hMSCs is the limited in vitro lifespan, but this can be overcome by introduction of the catalytic subunit of human telomerase reverse transcriptase (TERT). In this paper, the generation and characterization of a TERT-immortalized, non-tumorigenic human bone marrow-derived stromal mesenchymal model cell line is described. The resulting cell line, iMSC#3, maintained capacity to differentiate into osteoblasts and adipocytes, and growth characteristics comparable to primary hMSCs into after long-term culturing (155 population doublings). A detailed characterization of the mRNA- and microRNA transcriptomes during adipocyte differentiation also showed that the iMSC#3 recapitulates this process at the molecular level. Furthermore, iMSC#3 showed no major differences from hMSCs regarding surface marker expression. The cell line had a normal karyotype, and high-resolution array comparative genomic hybridization confirmed that the cells in general had normal copy number. The gene expression profiles of immortalized and primary hMSCs were also similar, whereas the corresponding methylation profiles were more diverse. In conclusion, a human bone marrow-derived stromal cell line was developed, which can be used for basic studies of mesenchymal cells and to functionally study transgenes that might play a role in oncogenic transformation.
2014-12-31 | GSE47647 | GEO
Project description:Human mesenchymal stromal cells (hMSCs) show great potential for clinical and experimental use. Self-renewal and multipotent differentiation capacity are key elements in the increasing interest of this cell type. However, a disadvantage of primary hMSCs is the limited in vitro lifespan, but this can be overcome by introduction of the catalytic subunit of human telomerase reverse transcriptase (TERT). In this paper, the generation and characterization of a TERT-immortalized, non-tumorigenic human bone marrow-derived stromal mesenchymal model cell line is described. The resulting cell line, iMSC#3, maintained capacity to differentiate into osteoblasts and adipocytes, and growth characteristics comparable to primary hMSCs into after long-term culturing (155 population doublings). A detailed characterization of the mRNA- and microRNA transcriptomes during adipocyte differentiation also showed that the iMSC#3 recapitulates this process at the molecular level. Furthermore, iMSC#3 showed no major differences from hMSCs regarding surface marker expression. The cell line had a normal karyotype, and high-resolution array comparative genomic hybridization confirmed that the cells in general had normal copy number. The gene expression profiles of immortalized and primary hMSCs were also similar, whereas the corresponding methylation profiles were more diverse. In conclusion, a human bone marrow-derived stromal cell line was developed, which can be used for basic studies of mesenchymal cells and to functionally study transgenes that might play a role in oncogenic transformation.
2014-12-31 | GSE47649 | GEO
Project description:Human mesenchymal stromal cells (hMSCs) show great potential for clinical and experimental use. Self-renewal and multipotent differentiation capacity are key elements in the increasing interest of this cell type. However, a disadvantage of primary hMSCs is the limited in vitro lifespan, but this can be overcome by introduction of the catalytic subunit of human telomerase reverse transcriptase (TERT). In this paper, the generation and characterization of a TERT-immortalized, non-tumorigenic human bone marrow-derived stromal mesenchymal model cell line is described. The resulting cell line, iMSC#3, maintained capacity to differentiate into osteoblasts and adipocytes, and growth characteristics comparable to primary hMSCs into after long-term culturing (155 population doublings). A detailed characterization of the mRNA- and microRNA transcriptomes during adipocyte differentiation also showed that the iMSC#3 recapitulates this process at the molecular level. Furthermore, iMSC#3 showed no major differences from hMSCs regarding surface marker expression. The cell line had a normal karyotype, and high-resolution array comparative genomic hybridization confirmed that the cells in general had normal copy number. The gene expression profiles of immortalized and primary hMSCs were also similar, whereas the corresponding methylation profiles were more diverse. In conclusion, a human bone marrow-derived stromal cell line was developed, which can be used for basic studies of mesenchymal cells and to functionally study transgenes that might play a role in oncogenic transformation.
2014-12-31 | GSE47648 | GEO
Project description:Human mesenchymal stromal cells (hMSCs) show great potential for clinical and experimental use due to their capacity to self-renew and differentiate into multiple mesenchymal lineages. However, disadvantages of primary cultures of hMSCs are the limited in vitro lifespan, and the variable properties of cells from different donors and over time in culture. In this article, we describe the generation of a telomerase-immortalized nontumorigenic human bone marrow-derived stromal mesenchymal cell line, and its detailed characterization after long-term culturing (up to 155 population doublings). The resulting cell line, iMSC#3, maintained a fibroblast-like phenotype comparable to early passages of primary hMSCs, and showed no major differences from hMSCs regarding surface marker expression. Furthermore, iMSC#3 had a normal karyotype, and high-resolution array comparative genomic hybridization confirmed normal copy numbers. The gene expression profiles of immortalized and primary hMSCs were also similar, whereas the corresponding DNA methylation profiles were more diverse. The cells also had proliferation characteristics comparable to primary hMSCs and maintained the capacity to differentiate into osteoblasts and adipocytes. A detailed characterization of the mRNA and microRNA transcriptomes during adipocyte differentiation also showed that the iMSC#3 recapitulates this process at the molecular level. In summary, the immortalized mesenchymal cells represent a valuable model system that can be used for studies of candidate genes and their role in differentiation or oncogenic transformation, and basic studies of mesenchymal biology.
| S-EPMC4172386 | biostudies-literature
Project description:Safety and Regenerative Properties of Immortalized Human Mesenchymal Stromal Cell Secretome
| PRJNA1312028 | ENA
Project description:This SuperSeries is composed of the SubSeries listed below.
2014-12-31 | GSE47651 | GEO
Project description:The secretome of mesenchymal stromal cells (MSCs) can efficiently stimulate regener-ation and therefore is a tempting remedy for “cell-free cellular therapy”. However, the usage of primary MSC cultures as secretome-producers for translation studies has ob-vious obstacles, including rapid aging of MSC cultures, the need for a large number of verified donors and donor-to-donor variability of secretome content. MSCs immortal-ization allows to overcome those limitations and to obtain secretome-producing cul-tures with prolonged lifetime. However, the efficacy and safety of such secretomes are critical issues, which limit their usage as therapeutic agents. In this study we have tested in large detail how the immortalization of MSC cultures affects the content, bi-ological activity and safety of their secretome. MSCs immortalization via overexpres-sion of human TERT gene does not significantly alter the qualitative and quantitative composition of their secretome or its activity according to the results of proteomic analysis, ELISA, qPCR and functional tests in vitro. Moreover, we have demonstrated that the secretome of immortalized MSCs does not contain detectable amounts of te-lomerase and does not possess any transforming activity. Altogether, our data suggest that immortalized MSC cultures may become a reliable source for obtaining standard-ized active secretome in large-scale quantities for clinical use.The secretome of mesenchymal stromal cells (MSCs) can efficiently stimulate regener-ation and therefore is a tempting remedy for “cell-free cellular therapy”. However, the usage of primary MSC cultures as secretome-producers for translation studies has ob-vious obstacles, including rapid aging of MSC cultures, the need for a large number of verified donors and donor-to-donor variability of secretome content. MSCs immortal-ization allows to overcome those limitations and to obtain secretome-producing cul-tures with prolonged lifetime. However, the efficacy and safety of such secretomes are critical issues, which limit their usage as therapeutic agents. In this study we have tested in large detail how the immortalization of MSC cultures affects the content, bi-ological activity and safety of their secretome. MSCs immortalization via overexpres-sion of human TERT gene does not significantly alter the qualitative and quantitative composition of their secretome or its activity according to the results of proteomic analysis, ELISA, qPCR and functional tests in vitro. Moreover, we have demonstrated that the secretome of immortalized MSCs does not contain detectable amounts of te-lomerase and does not possess any transforming activity. Altogether, our data suggest that immortalized MSC cultures may become a reliable source for obtaining standard-ized active secretome in large-scale quantities for clinical use.
2025-10-27 | PXD067843 | Pride
Project description:The secretome of mesenchymal stromal cells (MSCs) can efficiently stimulate regeneration and therefore is a tempting remedy for “cell-free cellular therapy”. However, the usage of primary MSC cultures as secretome-producers for translation studies has obvious obstacles, including rapid aging of MSC cultures, the need for a large number of verified donors and donor-to-donor variability of secretome content. MSCs immortalization allows to overcome those limitations and to obtain secretome-producing cultures with prolonged lifetime. However, the efficacy and safety of such secretomes are critical issues, which limit their usage as therapeutic agents. In this study we have tested in large detail how the immortalization of MSC cultures affects the content, biological activity and safety of their secretome. MSCs immortalization via overexpression of human TERT gene does not significantly alter the qualitative and quantitative composition of their secretome or its activity according to the results of proteomic analysis, ELISA, qPCR and functional tests in vitro. Moreover, we have demonstrated that the secretome of immortalized MSCs does not contain detectable amounts of telomerase and does not possess any transforming activity. Altogether, our data suggest that immortalized MSC cultures may become a reliable source for obtaining standardized active secretome in large-scale quantities for clinical use.
2025-08-28 | GSE306748 | GEO
Project description:Adipose-derived mesenchymal stromal cells (ADSC) are a promising source for cellular therapy of chronic wounds. However, the limited life span during in vitro expansion impedes their extensive use in clinical applications and basic research. We hypothesize that by introduction of an ectopic expression of telomerase into ADSC, the cells' lifespans could be significantly extended. To test this hypothesis, we aimed at engineering an immortalized human ADSC line using a lentiviral transduction with human telomerase (hTERT). ADSC were transduced with a third-generation lentiviral system and a hTERT codifying plasmid (pLV-hTERT-IRES-hygro). A population characterized by increased hTERT expression, extensive proliferative potential and remarkable (potent) multilineage differentiation capacity was selected. The properties for wound healing of this immortalized ADSC line were assessed after 17 passages. Their secretome induced the proliferation and migration of keratinocytes, dermal fibroblasts, and endothelial cells similarly to untransduced ADSC. Moreover, they sustained the complete re-epithelialization of a full thickness wound performed on a skin organotypic model. In summary, the engineered immortalized ADSC maintain the beneficial properties of parent cells and could represent a valuable and suitable tool for wound healing in particular, and for skin regenerative therapy in general.
| S-EPMC9408591 | biostudies-literature