Reprogramming of mouse bone marrow mesenchymal stem cells, mouse bone marrow mononuclear cells and mouse embryonic fibroblasts into induced pluripotent stem cells; a comparison of efficiencies / 第二军医大学学报

ABSTRACT

Objective To comparethe reprogramming efficiencies of mouse bone marrow mesenchymal stem cells (BMSCs), mouse bone marrow mononuclear cells (BM-MNCs) and mouse embryonic fibroblasts (MEFs) into induced pluripotent stem cells (iPS cells). Methods BMSCs, BM-MNCs and MEFs were infected with lentivirus (LV-efla-mOrt4- IRES-EGFP, LV-efla-mklf4-IRES-EGFP, LV-efla-mK/4-IRES-EGFP and LV-efla-mc-Mcc-IRES-EGFP) at a multiplicity of infection. Reprogramming efficiencies of BMSCs, BM-MNCs and MEFs were compared by counting the number of alkaline phosphatase (AP)-positive clones. Pluripotency of the clones was evaluated by detecting the expression of embryonic stem cells markers and assessing their ability to formembryoid bodies and teratomas. Results iPS cells derived from BMSCs, BM-MNCs and MEFs were all able to grow into clones with clear borders, to express enbryonic stem cell-specific cell surface markers (Nanog, Rex-1 and SSEA-1), and to express characteristic genes of all three germ layers both in vitro and vivo. The AP- positive clones derived from BMSCs were notably less than those from BM-MNCs and MEFs. Conclusion BMSCs, BM-MNCs and MEFs can all reprogram into iPS cells, but the reprogramming efficiency of BMSCs in adherent culture is lower than those of BM-MNCs and MEFs.