Optimized mouse ES cell culture system by suspension growth in a fully defined medium.
Mouse and human embryonic stem (mES and hES) cells have become one of the most intensively studied primary cell types in biomedical research. However, culturing ES cells is notoriously labor intensive. We have optimized current ES cell culture methods by growing mES cells in suspension in a defined medium. This protocol is unsurpassed in time efficiency and typically requires only 20 min of effective hands-on time per week. This protocol maintains a very high degree of pluripotent cells partly by mechanical separation of spontaneously differentiating cells. mES cells can be cultured for extended periods (>6 months) without the loss of pluripotency markers. High passage (>20) adherent mES cultures containing contaminating differentiated cells can be rescued and enriched in undifferentiated ES cells.
Andäng
Michael
M
Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm 171 77, Sweden.
Moliner
Annalena
A
Doege
Claudia A
CA
Ibañez
Carlos F
CF
Ernfors
Patrik
P
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods; D016895:Culture Media, Serum-Free / Q000032:analysis; D053595:Embryonic Stem Cells / Q000166:cytology*; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology
D016895:Culture Media, Serum-Free