Generation and Characterization of an immortalized human mesenchymal stromal cell line model [Illumina iMSC#3 and hMSC primary cultures]

ABSTRACT: 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE47648 | GEO | 2014/12/31

SECONDARY ACCESSION(S): PRJNA207292

REPOSITORIES: GEO