E-cadherin-coated plates maintain pluripotent ES cells without colony formation.
Embryonic stem (ES) cells cultured on gelatin-coated plates or feeder layers form tight aggregated colonies by the E-cadherin-mediated cell-cell adhesions. Here we show that murine ES cells do not make cell-cell contacts or form colonies when cultured on the plate coated with a fusion protein of E-cadherin and IgG Fc domain. The cells in culture retain all ES cell features including pluripotency to differentiate into cells of all three germ layers and germ-line transmission after extended culture. Furthermore, they show a higher proliferative ability, lower dependency on LIF, and higher transfection efficiency than colony-forming conditions. Our results suggest that aggregated colony formation might inhibit diffusion of soluble factors and increase cell-cell communication, which may result in a heterogeneous environment within and between surrounding cells of the colony. This method should enable more efficient and scalable culture of ES cells, an important step towards the clinical application of these cells.
Nagaoka
Masato
M
Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
Koshimizu
Uichi
U
Yuasa
Shinsuke
S
Hattori
Fumiyuki
F
Chen
Hao
H
Tanaka
Tomofumi
T
Okabe
Masaru
M
Fukuda
Keiichi
K
Akaike
Toshihiro
T
D000818:Animals; D015820:Cadherins / Q000502:physiology*; D002448:Cell Adhesion / Q000502:physiology; D002449:Cell Aggregation / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible; D003114:Colony-Forming Units Assay; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology* / Q000637:transplantation; D008297:Male; D051379:Mice; D008819:Mice, Nude; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology* / Q000637:transplantation; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction; D013724:Teratoma / Q000209:etiology; D013736:Testicular Neoplasms / Q000209:etiology; D014162:Transfection
D015820:Cadherins; D020099:Coated Materials, Biocompatible; D011993:Recombinant Fusion Proteins