抗凝固剤としてEDTA-2Kを用いたヒトの末梢血リンパ球の培養法
1983 年 18 巻 4 号 p. 319-326
詳細
1983 年 18 巻 4 号 p. 319-326
To use residual blood of blood examinations in radiation workers for chromosome examination, a culture method of EDTA-treated human peripheral lymphocytes by adding 3 metal rations (Ca, Zn, Fe) was developed.
EDTA-2K, an anticoagulant to be added to blood for blood examinations, is an effective inhibitor of PHA-induced lymphocyte transformation, whereas several metal cations have been reported to be effective in reversing EDTA inhibition of PHA-induced transformation. In the experiment, the solutions of 3 metal cations (Ca, Zn, Fe) in several concentrations were added to EDTA-treated blood for culture. It was found that when mol-concentrations of Ca, Zn, Fe ions were 3, 0.4, 0.25 times respectively as much as the concentration of EDTA in blood, a maximum number of mitotic cells could be observed.
In 50hr incubation, the number of mitotic cells observed, using the EDTA-2K method, was approximately 36% of that observed in PHA-stimulated culture of blood without EDTA using the conventional method. However, over a period of 74hr incubation, the results by the two methods became very similar.
