A focused microarray for screening rat embryonic stem cell lines.
Here, we describe a focused microarray for screening rat embryonic stem cells (ESCs) and provide validation data that this array can distinguish undifferentiated rat ESCs from rat trophoblast stem (TS) cells, rat extraembryonic endoderm cells, mouse embryonic fibroblast feeder cells, and differentiated rat ESCs. Using this tool, genuine rat ESC lines, which have been expanded in a conventional rat ESC medium containing two inhibitors (2i), for example, glycogen synthase kinase 3 (GSK3) and mitogen-activated protein kinase (MEK) inhibitors, and leukemia inhibitory factor, and genuine rat ESCs, which have been expanded in rat ESC medium containing four inhibitors (4i), for example, GSK3, MEK, Alk5, and Rho-associated kinase inhibitors were compared; as were genuine rat ESCs from 4 different strains of rats. Expression of Cdx2, a gene associated with trophoblast determination, was observed in genuine, undifferentiated rat ESCs from 4 strains and from both 2i and 4i ESC derivation medium. This finding is in contrast to undifferentiated mouse ESCs that do not express Cdx2. The rat ESC focused microarray described in this report has utility for rapid screening of rat ESCs. This tool will enable optimization of culture conditions in the future.
Department of Anatomy and Physiology, Kansas State University College of Veterinary Medicine, Manhattan, Kansas 66506, USA.
James
J
Hong
Hong
H
He
Phuoc
P
Bui
Ben
B
Ryba-White
Mohammad A K
MA
Rumi
Michael J
MJ
Soares
Debasree
D
Dutta
Soumen
S
Paul
Masaki
M
Kawamata
Takahiro
T
Ochiya
Qi-Long
QL
Ying
Pavan
P
Rajanahalli
Mark L
ML
Weiss
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000378:metabolism*; D061252:Feeder Cells; D005347:Fibroblasts / Q000378:metabolism; D020043:Genes, Essential; D005855:Germ Layers / Q000166:cytology; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D011916:Rats, Inbred F344; D020318:Rats, Long-Evans; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059467:Transcriptome; D014327:Trophoblasts / Q000166:cytology
D015415:Biomarkers