The Establishment And Committed Differentiation Towards Germ Cells Of Mouse Induced Pluripotent Stem Cell Lines

Induced pluripotent stem cells possessed the potentiality to differentiate into all kinds of tissues and cells, and this technique could circumvent the ethical conflict and avoid immune rejection after transplantation which should be faced in the application of embryonic stem cells. The establishment and committed differentiation of iPSC have become one of the hot research fields nowadays. As we all know, germ cells play an important role in ontogeny. The germ cells, obtained from iPS differentiation in vitro, could not only be utilized as gene-modified cells, but the transgenic animals could be acquired after fertilization. Meanwhile, they could be used in the treatment of infertility and aphoria.In this study, Yamanaka OSKM, the four transcription factors were transfered into mice embyyonic fibroblasts by pMXs-retroviral vector, combined with Tet-on inducible system controled by Dox. And then mice iPS lines were successfully established and their biological characteristics were analyzed and identified. The experiments of AP activity, the number of chromosome, stem cell markers, and teratomas in vivo and in vitro differentiation has showed that the iPS cells had the similar characteristics to the ES cells.The EBs were formed, when the established mouse iPS cells were cultured in a hanging drop at3x104cells/mL concentration. And during the differentiated culture of the EBs, the continuing beat phenomenon could be observed. The immunofluorescence staining showed that the cardiac-specific protein GATA4was expressed in the differentiated cells.A certain concentration of RA-induced medium could promote mouse iPS cells to differentiate into germ cell precursor cells, which expressed specific proteins DAZL. All these lay the technical foundation for further obtaining the functional germ cells in vitro.