Abstract
2-Mercaptoethanol (2-ME) is used as a medium supplement to enhance the proliferation of lymphocytes culturedin vitro. In this study, we have examined the effects of 2-ME on cell growth and on SCE induction in cultures of unstimulated and phytohemagglutinin (PHA)-stimulated Fischer 344 rat lymphocytes. There were virtually no metaphases detected in cells cultured without PHA. In PHA-stimulated cultures, 2-ME decreased SCE-frequency but it enhanced SCE frequency in the presence of S to 12.5 µM bromodeoxyuridine (BRd U). Both mitotic and replication indices were increased in the PHA/2-ME system. The levels of incorporated exogenous thymidine, in the presence of 2-ME, were relatively low in unstimulated cells, suggesting that 2-ME is not mitogenic for T-cells. However, 2-ME enhanced PHA-induced response of T-cells as evidenced by increased levels of thymidine incorporation into cellular DNA. The growth promoting effects and the decrease in SCE frequency caused by 2-ME upon PHA stimulation indicate that 2-ME may alter the nature of interaction between PHA and cellular activating properties or the replicative processes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- BRdU:
-
bromodeoxyuridine
- FBS:
-
fetal bovine serum
- SCE:
-
sister-chromatid exchanges
- HEPES:
-
N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid
- IL-2:
-
interleukin-2
- 2-ME:
-
2-mercaptoethanol
- PBS:
-
phosphate buffered saline
- PHA:
-
phytohemagglutinin
- MI:
-
mitotic index
- RI:
-
replication index
- NADH:
-
nicotinamide adenine dinucleotide (reduced form)
References
BROGGER, A. and WAKSVIK, H. (1978). Further evidence that the chromatid gap is a folding defect. Hereditas 89:131–132.
BROOME, J.D., and JENG, M.W. (1973). Promotion of replication in lymphoid cells by specific thiols and disulfidesin vitro. Effects on mouse lymphoma cells in comparison with splenic lymphocytes. J. Exp. Med. 138:574–592.
BUDROE, J.D., SHADDOCK, J.G. and CASCIANO, D.A. (1984). A study of the potential genotoxicity of methapyrilene and related antihistamines using the hepatocyte/DNA repair assay. Mutation Res.135:131–137.
CHANG, M.P., STREHLER, B.L. and MAKINODAN, T. (1982). Requirements of 2-mercaptoethanol forin vitro growth factor production by T-cells and vulnerability of the response to age. Mech. Age. Develop. 20:65–73.
CLICK, R.E., BENCK, L. and ALTER, B.J. (1972). Enhancement of antibody synthesisin vitro by mercaptoethanol. Cell Immunol.3:156–160.
FANGER, M.W., HART, D.A. and WELLS, J.V. (1970). Enhancement of reducing agents of the transformation of human and rabbit peripheral lymphocytes. J. Immunol.105:1043–1045.
GOON, D. and CONNER, M.K. (1984). Simultaneous assessment of ethyl carbamate-induced SCEs in murine lymphocytes, bone marrow and alveolar macrophage cells. Carcinogenesis5(3):399–402.
HEARTLEIN, M.W., O'NEILL, J.P. and PRESTON, R.J. (1983). SCE induction is proportional to substitution in DNA for thymidine by C1dU and BrdU. Mutation Res.107:103–109.
KLIGERMAN, A.D., WILMER, J.L. and EREXSON, G.L. (1981). Lymphocyte culture system for assessing sister chromatid exchange afterin vivo exposure to genotoxic agents. Environ. Mutagen.3:531–541.
KLIGERMAN, A.D., EREXSON, G.L. and WILMER, J.L. (1984). Development of a rodent peripheral blood lymphocyte culture system to detect cytogenetic damagein vivo. In Sister-chromatid exchanges: Twenty-five Years of Experimental Research, R.R. Tice and A.E. Hollaender (eds.), pp. 569–584. Plenum, New York.
LINDBLAD, A. and LAMBERT, B. (1981). Relation between sister chromatid exchange, cell proliferation and proportion of B and T cells in human lymphocyte cultures. Hum. Genet. 57:31–34.
RICHER, C.L., SENECAL-QUEVILLON, M. and DOQUETTE, P. (1987). Analysis of low concentrations of 5-bromo-2-deoxyuridine on sister chromatid exchanges in human lymphocytes. Genome29:165–168.
ROSIN, C.E. and STICH, H.F. (1978). Inhibitory effect of reducing agents on N-acetoxy and N-hydroxy-2-acetylaminofluorene-induced mutagenesis. Cancer Res. 38:1307–1310.
SASAKI, N.S. and MATSUBARA, S. (1977). Free radical scavenging in protection of human lymphocytes against chromosome aberration formation by gamma-ray irradiation. Intl. J. Rad. Biol.32:439–445.
SCHNEIDER, E.L. and LEWIS, J. (1981). Aging and sister chromatid exchange induction and cell cycle kinetics in Ehrlich ascites tumor cells. Mech. Age. Develop.17:327–330.
SPEIT, G., WOLF, M. and VOGEL, W. (1980). The effect of sulfhydryl compounds on sister chromatid exchanges. Mutation Res. 78:267–272.
STOBO, J.D., ROSENTHAL, A.S. and PAUL, W.E. (1972). Functional heterogeneity of murine lymphoid cells. I. Responsiveness to and surface binding of concanavalin A and phytohemagglutinin. J. Immunol.108:1–17.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Aidoo, A., Morris, S.M., Domon, O.E. et al. Modulation of SCE induction and cell proliferation by 2-mercaptoethanol in phytohemagglutinin-stimulated rat lymphocytes. Cell Biol Toxicol 5, 237–248 (1989). https://doi.org/10.1007/BF01795353
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01795353