title: Basal medium composition and serum or serum replacement concentration influences on the maintenance of murine embryonic stem cells.
abstract: The expansion of stem cell numbers while retaining their developmental properties is a bioprocess challenge. We compared the growth rates and embryoid body (EB) formation yields of R1 and EFC murine embryonic stem cells (mESC) cultured in two basal media (DMEM or DMEM:F12) with additions of 1.7-15% fetal bovine serum (FBS) or serum replacer (KOSR). Whereas the basal medium or KOSR dose did not have a significant effect on growth rate for either cell line, increasing doses of KOSR had a significant negative effect on the EB yield of EFC cells. Use of DMEM:F12 and increasing doses of FBS independently and significantly increased the growth rate for both cell lines. DMEM:F12 also significantly increased EB yields for both cell lines. The results show that use of DMEM:F12 and several-fold lower than conventional concentrations of KOSR can efficiently support maintenance of mESC and that KOSR should be dose as well as lot optimized.
authors.affiliation: Michael Smith Laboratories, University of British Columbia, 2185 East Mall, Vancouver, BC, V6T 1Z4, Canada.
authors.forename: Muhammad A
authors.initials: MA
authors.lastname: Chaudhry
authors.forename: Timothy Z
authors.initials: TZ
authors.lastname: Vitalis
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Bowen
authors.forename: James M
authors.initials: JM
authors.lastname: Piret