Abstract
Two purified serum protein fractions, fetuin and serum albumin, will replace whole or dialyzed serum in supporting the growth of single S3 HeLa cells in an otherwise chemically defined nutrient solution. In the serum-free medium, single S3 cells will form macroscopic colonies with essentially 100 per cent efficiency. The generation time of S3 cells in the serum-free medium is approximately 50 per cent greater than that observed in an optimal, serum-containing medium. All components of the serum-free medium are available commercially, except fetuin, which can easily be prepared in substantial quantities. The problem of the purity of the protein preparations and of their possible roles in promoting cell growth is discussed.
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Selected References
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- CHU E. H., GILES N. G. Comparative chromosomal studies on mammalian cells in culture. I. The HeLa strain and its mutant clonal derivatives. J Natl Cancer Inst. 1958 Feb;20(2):383–401. [PubMed] [Google Scholar]
- DEUTSCH H. F. Fetuin: the mucoprotein of fetal calf serum. J Biol Chem. 1954 Jun;208(2):669–678. [PubMed] [Google Scholar]
- Fisher H. W., Puck T. T., Sato G. MOLECULAR GROWTH REQUIREMENTS OF SINGLE MAMMALIAN CELLS: THE ACTION OF FETUIN IN PROMOTING CELL ATTACHMENT TO GLASS. Proc Natl Acad Sci U S A. 1958 Jan;44(1):4–10. doi: 10.1073/pnas.44.1.4. [DOI] [PMC free article] [PubMed] [Google Scholar]
- HJERTEN S., LEVIN O., TISELIUS A. Protein chromatography on calcium phosphate columns. Arch Biochem Biophys. 1956 Nov;65(1):132–155. doi: 10.1016/0003-9861(56)90183-7. [DOI] [PubMed] [Google Scholar]
- JACQUEZ J. A., BARRY E. Tissue culture media; the essential non-dialyzable factors in human placental cord serum. J Gen Physiol. 1951 Jul;34(6):765–774. doi: 10.1085/jgp.34.6.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LIEBERMAN I., OVE P. A protein growth factor for mammalian cells in culture. J Biol Chem. 1958 Sep;233(3):637–642. [PubMed] [Google Scholar]
- MEYERS W. M., DEUTSCH H. F. Immunological studies of fetuin. Arch Biochem Biophys. 1955 Jan;54(1):38–44. doi: 10.1016/0003-9861(55)90006-0. [DOI] [PubMed] [Google Scholar]
- NEUMAN R. E., McCOY T. A. Growth-promoting properties of pyruvate oxal-acetate, and alpha-ketoglutarate for isolated Walker carcinosarcoma 256 cells. Proc Soc Exp Biol Med. 1958 Jun;98(2):303–306. doi: 10.3181/00379727-98-24025. [DOI] [PubMed] [Google Scholar]
- PUCK T. T., CIECIURA S. J., ROBINSON A. Genetics of somatic mammalian cells. III. Long-term cultivation of euploid cells from human and animal subjects. J Exp Med. 1958 Dec 1;108(6):945–956. doi: 10.1084/jem.108.6.945. [DOI] [PMC free article] [PubMed] [Google Scholar]
- PUCK T. T., MARCUS P. I., CIECIURA S. J. Clonal growth of mammalian cells in vitro; growth characteristics of colonies from single HeLa cells with and without a feeder layer. J Exp Med. 1956 Feb 1;103(2):273–283. doi: 10.1084/jem.103.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sato G., Fisher H. W., Puck T. T. Molecular Growth Requirements of Single Mammalian Cells. Science. 1957 Nov 8;126(3280):961–964. doi: 10.1126/science.126.3280.961. [DOI] [PubMed] [Google Scholar]
- TJIO J. H., PUCK T. T. Genetics of somatic mammalian cells. II. Chromosomal constitution of cells in tissue culture. J Exp Med. 1958 Aug 1;108(2):259–268. doi: 10.1084/jem.108.2.259. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WAYMOUTH C. Simple nutrient solutions for animal cells. Tex Rep Biol Med. 1955;13(3):522–536. [PubMed] [Google Scholar]
- WINZLER R. J. Determination of serum glycoproteins. Methods Biochem Anal. 1955;2:279–311. doi: 10.1002/9780470110188.ch10. [DOI] [PubMed] [Google Scholar]