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  • Title: [Culture of mouse embryo for the toxicological evaluation of drugs in vitro. (2). Differential staining of sister chromatids of blastocyst cell].
    Author: Kamata K, Oh-Ishi S.
    Journal: Nihon Yakurigaku Zasshi; 1983 Dec; 82(6):419-27. PubMed ID: 6199269.
    Abstract:
    A method for differential staining of sister chromatids of cell from mouse blastocyst was devised. Mouse embryo was cultured for 24 hr in BMOC-III medium from the 8 cell stage and then transferred into the medium containing 5-bromodeoxyuridine (BrdU) for a further 24 hr. At 4 hr prior to the termination of culture, colchicine was added to arrest the cell division in metaphase. Cells were fixed on a slide glass, air-dried, stained with Hoechst 33258, lighted with a mercury light, and stained with Giemsa. Basic conditions for the differential staining such as the concentration of BrdU and lighting were examined. When the mercury light (400 W) was used at 15 cm distance for 60-90 min, sister chromatids could be clearly differentiated with a BrdU concentration of 3-30 ng/ml. Under these concentrations of BrdU, the frequency of sister chromatid exchange (SCE) was almost the same as the control level. Thus the concentration could not affect the SCE frequency. This method can be used for the evaluation of drug toxicity of genetic or mutational effects on mammalian embryos.
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