Single cell derived murine embryonic stem cell clones stably express Rex1-specific green fluorescent protein and their differentiation study.
Embryonic stem cells (ESCs) often display high rates of apoptosis and spontaneous differentiation in routine culture, thus bring the proliferation of these cells highly inefficient. Moreover, little is known about the factors that are indispensable for sustaining self-renewal. To surmount these issues, we established transgenic mES cell lines expressing the enhanced green fluorescent protein (EGFP) under the control of the Rex1 promoter which is a key regulator of pluripotency in ES cells. In addition, we provided a simplified and improved protocol to derive transgenic mESCs from single cell. Finally, we showed that embryoid body (EB) development was faster than adherent differentiation in terms of differentiation ratio by real-time tracking of the EGFP expression. Therefore, these cell lines can be tracked and selected both in vitro and in vivo and should be invaluable for studying the factors that are indispensable for maintaining pluripotency.
Chen
Xiaopan
X
College of Life Sciences, Zhejiang University, No. 338, Yu-Hang-Tang Road, Hangzhou, Zhejiang 310058, China.
Wu
Rongrong
R
Feng
Shumei
S
Gu
Bin
B
Dai
Licheng
L
Zhang
Ming
M
Zhao
Xiaoli
X
D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D002999:Clone Cells; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D007621:Karyotyping; D051379:Mice; D008819:Mice, Nude; D016513:Mice, SCID; D008856:Microscopy, Fluorescence / Q000379:methods; D009374:Neoplasms, Experimental / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D011401:Promoter Regions, Genetic / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013724:Teratoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
C513672:Zfp42 protein, mouse; D014157:Transcription Factors; D049452:Green Fluorescent Proteins