Lymphocyte system, cultured

Various types of cells can be scored for chromosomal aberrations. Spermatogonial cells, spermatocytes, oocytes, early embryos, bone marrow cells, and lymphocytes have been used successfully after in vivo treatments of an animal. For short-term cytogenetic analysis, however, an in vitro cell-culture assay is often chosen. Such in vitro assays usually use a peripheral lymphocyte system or a monolayer cell-culture system. 

Results of methyl parathion assays involving effects on chromosomes have also been contradictory. For sister chromatid exchange, Waters et al. (1982) reported a positive response in Chinese hamster ovary cells only in the presence of metabolic activation system, while methyl parathion tested positive without a metabolic activation system in Chinese hamster V79 cells (Chen et al. 1981), cultured normal human lymphoid cells (Chen et al. 1981 Gomez-Arroyo et al. 1987 Sobti et al. 1982), and Burkitt s l5miphoma cells (Chen et al. 1981). Chen et al. (1981) found a significant dose-related increase in sister chromatid exchange in both hamster and human cultured cells, but dose-related cell cycle delays were less pronounced in human cell lines than in V79 cells. Negative results were obtained for chromosomal aberrations in human lymphocytes without a metabolic activation system (Kumar et al. 1993). 

Jyonouchi, H. et al., Studies of immunomodulating actions of carotenoids. I. Effects of P-carotene and astaxanthin on murine lymphocyte functions and cell surface marker expression in in vitro culture system, Nutr. Cancer, 19, 93, 1991. 

Established cell lines, cell strains or primary cell cultures may be used. The most often used are Chinese hamster cell lines and human peripheral blood lymphocytes. The merits of these two cell lines have been reported (Ishidate and Hamois, 1987 Kirkland and Gamer, 1987). The cell system must be validated and consistently sensitive to known clastogens. 

Mammalian cells in vitro are exposed to the test chemical with and without an exogenous mammalian metaboUc activation system and cultured for two rounds of replication in bromodeox3Uiridine (BrdU) containing medium. After treatment with a spindle inhibitor (e.g., colchicine) to accumulate cells in a metaphase-Uke stage of mitosis (c-metaphase), cells are harvested, stained, and metaphase cells analyzed for SCEs. Primary cultures (e.g., human lymphocytes) or established cell lines (e.g., Chinese hamster ovary or lung cells) may be used in the assay. At least three adequately spaced concentrations of the test substance should be used. 

A Cell Culture System That Mimics Chronic Lymphocytic Leukemia Cells Microenvironment for Drug Screening and Characterization... 

Striking clinical results have been reported in inflammatory and auto-immune diseases, although in a somewhat limited number of cases . A low molecular mass chromosome-breaking agent was identified in the serum of patients with systemic lupus erythematosus. The chromosome aberrations in cultures of normal lymphocytes in the presence of this factor were reduced to normal values by the addition of BESOD to the culture medium... 

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