Stem Cells and Development 2014-10-01

Generation and characterization of an immortalized human mesenchymal stromal cell line.

Magne Skårn, Paul Noordhuis, Meng-Yu Wang, Marjan Veuger, Stine Henrichson Kresse, Eivind Valen Egeland, Francesca Micci, Heidi Maria Namløs, Anne-Mari Håkelien, Solveig Mjelstad Olafsrud, Susanne Lorenz, Guttorm Haraldsen, Gunnar Kvalheim, Leonardo Andrés Meza-Zepeda, Ola Myklebost

Index: Stem Cells Dev. 23(19) , 2377-89, (2014)

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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.