Generation of transgenic rats through induced pluripotent stem cells.
The rat is an important animal model for human disease research. Using inhibitors of glycogen synthase kinase 3 and MAPK signaling pathways, rat embryonic stem cells and rat induced pluripotent stem cells (riPSCs) have been derived. However, unlike rat embryonic stem cells, germ line competent riPSCs have only been derived from Wistar rats at low efficiency. Here, we found that an optimized induction medium containing knock-out serum replacement and vitamin C improved the rate and efficiency of riPSCs generation from Dark Agouti rat fibroblasts and Sertoli cells. riPSCs maintained an undifferentiated status for >30 passages and could differentiate into various cells types including germ cells when injected into rat blastocysts. Moreover, transgenic riPSCs could be generated through the PiggyBac transposon, which could be used to generate transgenic rats through germ line transmission. riPSCs can be used as a novel tool in genetic and genomic studies of the rat.
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China; Key Laboratory of Translational Stem Cell Research, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
Ming-Gui
MG
Jiang
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Key Laboratory of Translational Stem Cell Research, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
Tianda
T
Li
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Key Laboratory of Translational Stem Cell Research, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Graduate School of the Chinese Academy of Sciences, Beijing 100039, China.
Chunjing
C
Feng
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Key Laboratory of Translational Stem Cell Research, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
Rui
R
Fu
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing, Jiangsu 210000, China.
Yan
Y
Yuan
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing, Jiangsu 210000, China.
Quan
Q
Zhou
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; College of Life Sciences, Northeast Agricultural University of China, Harbin 150030, China.
Xin
X
Li
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Key Laboratory of Translational Stem Cell Research, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
Haifeng
H
Wan
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Key Laboratory of Translational Stem Cell Research, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
Liu
L
Wang
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Key Laboratory of Translational Stem Cell Research, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
Wei
W
Li
College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China.
Yamei
Y
Xiao
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Key Laboratory of Translational Stem Cell Research, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China. Electronic address: xyzhao@ioz.ac.cn.
Xiao-Yang
XY
Zhao
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Key Laboratory of Translational Stem Cell Research, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China. Electronic address: qzhou@ioz.ac.cn.
Qi
Q
Zhou
D000818:Animals; D001755:Blastocyst; D005260:Female; D005347:Fibroblasts; D006801:Humans; D057026:Induced Pluripotent Stem Cells; D008297:Male; D051379:Mice; D051381:Rats; D011916:Rats, Inbred F344; D017207:Rats, Sprague-Dawley; D055647:Rats, Transgenic; D017208:Rats, Wistar; D012708:Sertoli Cells
Induced Pluripotent Stem Cells; Rat; Reprogramming; Transgenic; Vitamin C