Abstract
Suspension cultures of Chinese hamster cells (line CHO), which had stopped dividing and were arrested in G1 following growth to high cell concentrations in F-10 medium, could be induced to reinitiate DNA synthesis and to divide in synchrony upon addition of the appropriate amounts of isoleucine and glutamine. Both amino acids were required to initiate resumption of cell-cycle traverse. Deficiencies in other amino acids contained in F-10 medium did not result in accumulation of cells in G1, indicating a specific action produced by limiting quantities of isoleucine and glutamine. In the presence of sufficient glutamine, approximately 2 x 10-6 M isoleucine was required for all cells to initiate DNA synthesis in a population initially containing 1.5 x 105 cells/ml. Under similar conditions, about 4 x 10-6 M isoleucine was required for all G1-arrested cells to progress through cell division. In contrast, 1 x 10-4 M glutamine was necessary for maximum initiation of DNA synthesis in G1 cells, along with sufficient isoleucine. A technique for rapid production of G1-arrested cells is described in which cells from an exponentially growing population placed in F-10 medium deficient in both isoleucine and glutamine or isoleucine alone accumulated in G1 after 30 hr.
Full Text
The Full Text of this article is available as a PDF (438.7 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- CHANOCK R. M., HAYFLICK L., BARILE M. F. Growth on artificial medium of an agent associated with atypical pneumonia and its identification as a PPLO. Proc Natl Acad Sci U S A. 1962 Jan 15;48:41–49. doi: 10.1073/pnas.48.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Griffiths J. P., Pirt S. J. The uptake of amino acids by mouse cells (strain LS) during growth in batch culture and chemostat culture: the influence of cell growth rate. Proc R Soc Lond B Biol Sci. 1967 Nov 14;168(1013):421–438. doi: 10.1098/rspb.1967.0073. [DOI] [PubMed] [Google Scholar]
- HAM R. G. An improved nutrient solution for diploid Chinese hamster and human cell lines. Exp Cell Res. 1963 Feb;29:515–526. doi: 10.1016/s0014-4827(63)80014-2. [DOI] [PubMed] [Google Scholar]
- LITTLEFIELD J. W. DNA synthesis in partially synchronized L cells. Exp Cell Res. 1962 Mar;26:318–326. doi: 10.1016/0014-4827(62)90184-2. [DOI] [PubMed] [Google Scholar]
- MOHBERG J., JOHNSON M. J. AMINO ACID UTILIZATION BY 929-L FIBROBLASTS IN CHEMICALLY DEFINED MEDIA. J Natl Cancer Inst. 1963 Sep;31:611–625. [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]
- Tobey R. A., Ley K. D. Regulation of initiation of DNA synthesis in Chinese hamster cells. I. Production of stable, reversible G1-arrested populations in suspension culture. J Cell Biol. 1970 Jul;46(1):151–157. doi: 10.1083/jcb.46.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]