Efficient derivation of embryonic stem cells from NOD-scid Il2rg (-/-) mice.

Kang Liu,Haibo Li,Jinhua Wen,Weifeng Yang,Zhenchuan Miao,Riguo Fang,Hongkui Deng

doi:10.1007/s13238-015-0209-6

Abstract

Efficient derivation of embryonic stem cells from NOD-scid Il2rg (-/-) mice.

Highlights

The stem cell field has pursued the development of humanized mice to further enhance our understanding of human hematopoiesis, innate immunity, infectious diseases, cancer biology and regenerative medicine
To derive non-obese diabetic (NOD)-scid interleukin 2 receptor gamma chain (Il2rg)−/− embryonic stem cell lines, a total of 28 blastocysts were recovered from NOD-scid Il2rg−/− mice (NPGTM, VITALSTAR) and cultured on MEF feeder cells under N2/B27 medium supplemented with the 2i/LIF, which was previously reported for use in harvesting and culturing rodent pluripotent stem cells (Ying et al, 2008)
All 14 ES cell lines showed dome-shaped colonies with smooth boundaries (Fig. 1A) and we chose 3 ES cell lines for further analysis. These NPGESCs had normal karyotypes (40, XY for male) and maintained alkaline phosphatase (AP) activity for more than 15 passages (Fig. 1A and 1F). They grew with a doubling time similar to those of TT2 mouse embryonic stem cells and chemically induced pluripotent stem cells (CiPSCs) that have been established previously (Hou et al, 2013) (Fig. 1E)

Summary

Introduction

The stem cell field has pursued the development of humanized mice to further enhance our understanding of human hematopoiesis, innate immunity, infectious diseases, cancer biology and regenerative medicine. To derive NOD-scid Il2rg−/− embryonic stem cell lines, a total of 28 blastocysts were recovered from NOD-scid Il2rg−/− mice (NPGTM, VITALSTAR) and cultured on MEF (mouse embryonic fibroblasts) feeder cells under N2/B27 medium supplemented with the 2i/LIF, which was previously reported for use in harvesting and culturing rodent pluripotent stem cells (Ying et al, 2008). We successfully established 14 ES cell lines from the 28 blastocysts (Fig. 1B).

Results

Conclusion