title: Generation and characterization of an immortalized human mesenchymal stromal cell line.
abstract: 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.
authors.affiliation: 1 Department of Tumor Biology, Institute of Cancer Research, The Norwegian Radium Hospital, Oslo University Hospital , Oslo, Norway .
authors.forename: Magne
authors.initials: M
authors.lastname: Skårn
authors.forename: Paul
authors.initials: P
authors.lastname: Noordhuis
authors.forename: Meng-Yu
authors.initials: MY
authors.lastname: Wang
authors.forename: Marjan
authors.initials: M
authors.lastname: Veuger
authors.forename: Stine Henrichson
authors.initials: SH
authors.lastname: Kresse
authors.forename: Eivind Valen
authors.initials: EV
authors.lastname: Egeland
authors.forename: Francesca
authors.initials: F
authors.lastname: Micci
authors.forename: Heidi Maria
authors.initials: HM
authors.lastname: Namløs
authors.forename: Anne-Mari
authors.initials: AM
authors.lastname: Håkelien
authors.forename: Solveig Mjelstad
authors.initials: SM
authors.lastname: Olafsrud
authors.forename: Susanne
authors.initials: S
authors.lastname: Lorenz
authors.forename: Guttorm
authors.initials: G
authors.lastname: Haraldsen
authors.forename: Gunnar
authors.initials: G
authors.lastname: Kvalheim
authors.forename: Leonardo Andrés
authors.initials: LA
authors.lastname: Meza-Zepeda
authors.forename: Ola
authors.initials: O
authors.lastname: Myklebost
mesh_terms: D017667:Adipocytes / Q000166:cytology*; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D049109:Cell Proliferation / Q000502:physiology*; D002471:Cell Transformation, Neoplastic / Q000378:metabolism; D055028:Comparative Genomic Hybridization; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*