E-cadherin gene-engineered feeder systems for supporting undifferentiated growth of mouse embryonic stem cells.
Conventionally, embryonic stem (ES) cells are cultured on a cell layer of mouse embryonic fibroblasts (MEFs) as feeder cells to support undifferentiated growth of ES cells. In this study, cell-cell interactions between mouse ES and feeder cells were artificially engineered via an epithelial cell adhesion molecule, E-cadherin, whose expression is considerable in ES cells. Mouse mesenchymal STO and NIH3T3 cells that were genetically engineered to express E-cadherin were used in ES cell cultures as feeder cells. ES cells cultured on the E-cadherin-expressing feeder cells maintained the expression of stem cell markers, alkaline phosphatase (AP), Oct3/4, Nanog and Sox2, and the efficiency of AP-positive colony formation was comparable to MEFs, and much better than parental STO and NIH3T3 cells. Furthermore, ES cells maintained on the E-cadherin-expressing feeder cells possessed the ability to differentiate into the three germ layers both in vitro and in vivo. The results indicated that E-cadherin expression in feeder cells could improve the performance of feeder cells, which may be further applicable to create new artificial feeder cell lines.
Horie
Masanobu
M
Department of Chemical Engineering, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.
Ito
Akira
A
Kiyohara
Takehiko
T
Kawabe
Yoshinori
Y
Kamihira
Masamichi
M
D000818:Animals; D001483:Base Sequence; D015820:Cadherins / Q000235:genetics*; D002450:Cell Communication; D002454:Cell Differentiation; D049109:Cell Proliferation; D018920:Coculture Techniques / Q000379:methods*; D003114:Colony-Forming Units Assay; D017931:DNA Primers / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D015870:Gene Expression; D005818:Genetic Engineering; D051379:Mice; D041681:NIH 3T3 Cells; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D015398:Signal Transduction
D015820:Cadherins; D017931:DNA Primers