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.
Similar content being viewed by others
References
Dulbecco R, Freeman G (1959) Plaque production by the polyoma virus. Virology 8(3):396–397. doi:10.1016/0042-6822(59)90043-1
Eagle H (1955) Nutrition needs of mammalian cells in tissue culture. Science 122(3168):501–514. doi:10.1126/science.122.3168.501
Keller G, Kennedy M, Papayannopoulou T, Wiles MV (1993) Hematopoietic commitment during embryonic stem cell differentiation in culture. Mol Cell Biol 13(1):473–486
Mather JP, Sato GH (1979) The growth of mouse melanoma cells in hormone-supplemented, serum-free medium. Exp Cell Res 120(1):191–200. doi:10.1016/0014-4827(79)90549-4
Nagy A, Rossant J, Nagy R, Abramownewerly W, Roder JC (1993) Derivation of completely cell culture-derived mice from early-passage embryonic stem-cells. Proc Natl Acad Sci USA 90(18):8424–8428. doi:10.1073/pnas.90.18.8424
Nichols J, Evans EP, Smith AG (1990) Establishment of germ-line-competent embryonic stem (ES) cells using differentiation inhibiting activity. Development 110(4):1341–1348
Palmqvist L, Glover CH, Hsu L, Lu M, Bossen B, Piret JM, Humphries RK, Helgason CD (2005) Correlation of murine embryonic stem cell gene expression profiles with functional measures of pluripotency. Stem Cells 23(5):663–680. doi:10.1634/stemcells.2004-0157
Price PJ, Goldsborough MD, Tilkins ML (1998) Embryonic stem cell serum replacement. International Patent Application WO98/30679
Schmelzer AE, de Zengotita VM, Miller WM (2000) Considerations for osmolality measurement under elevated pCO(2): comparison of vapor pressure and freezing point osmometry. Biotechnol Bioeng 67(2):189–196. doi:10.1002/(SICI)1097-0290(20000120)67:2<189::AID-BIT8>3.0.CO;2-U
Acknowledgments
The authors gratefully acknowledge Drs. Connie Eaves and Michael O’Connor for their helpful discussions. Financial support for this work was provided by StemCell Technologies, Inc. (Vancouver, BC) and the Canadian Stem Cell Network of Centres of Excellence.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Chaudhry, M.A., Vitalis, T.Z., Bowen, B.D. et al. Basal medium composition and serum or serum replacement concentration influences on the maintenance of murine embryonic stem cells. Cytotechnology 58, 173–179 (2008). https://doi.org/10.1007/s10616-008-9177-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10616-008-9177-5