Abstract:

Objective To establish a cell bank of mouse induced pluripotent stem cells (iPSCs), which would be a useful resource for the research of stem cells and iPSCs. Methods C57BL/6J mouse embryonic fibroblasts were transfected with lentivirus with Sox2, Oct4, Klf4 and c-Myc. Twelve days after viral infection, mouse iPSCs clones were picked and expanded, the alkaline phosphatase (AP) was stained. The iPSCs were hanging cultured in vitro and subcutaneously inoculated into BABL/c nude mice. All-trans retinoic acid (RA) was introduced to the media to induce smooth muscle cell (SMC) differentiation of iPSCs, and SMC-specific gene (smMHC and SMA) expression were measured by RT-PCR analyses. PCR method was employed for species identification and detection of mycoplasma. Results Twelve AP positive mouse iPSCs clonal strains were picked and expanded, and 9 of them formed teratomas in BALB/c nude mice. HE staining of the teratoma showed various types of tissues. RT-PCR of iPSCs in RA media showed positive expression of smMHC and SMA. The iPSCs clonal strains were identified as murine species, without mycoplasma contamination was found, and were deposited by cell resource center. Conclusion A model of mouse iPSCs was successfully established, which may provide available resources for reprogramming and directed differentiation mechanism research of ES cells and iPSCs.

Key words: Induced pluripotent stem cells, Viral infection, AP staining, RT-PCR, Mouse