Influence of anticoagulants and storage temperatures on blood counts and mitotic index of blood samples collected for cytogenetic biodosimetry.

Research paper by Yohei Y Fujishima, Syuki S Kanahama, Shigeki S Hagino, Shiori S Natsubori, Hitoshi H Saito, Ayaka A Azumaya, Kentaro K Ariyoshi, Akifumi A Nakata, Kosuke K Kasai, Kyogo K Yamada, Yasushi Y Mariya, Mitsuaki A MA Yoshida, Tomisato T Miura

Indexed on: 31 Oct '18Published on: 31 Oct '18Published in: International journal of radiation biology


In order to establish suitable protocols of blood culture to obtain sufficient numbers of metaphases for dicentric chromosome assay (DCA), we have examined the effect of storage temperature, storage time, and anticoagulant type. Peripheral blood was collected from five healthy donors with lithium heparin and ethylenediaminetetraacetic acid dipotassium salt (EDTA-2K). These samples were irradiated with X-rays at 3 Gy or sham; the samples were further divided into groups that were either stored at room temperature (RT), or 5.2 ± 1.0 °C. After 6, 24, 48, 72, and 168 h of storage, both blood counts and the mitotic index (MI) were analyzed. Heparinized blood samples stored under cold conditions exhibited low white blood cell, lymphocyte, and platelet counts. EDTA-treated blood samples did not show such obvious changes in cell counts. After 6 h of storage, heparinized blood samples stored at RT had MI of 21.5-29.3%. Similar MI was obtained in the EDTA-washed group stored for 6, 24, 48, and 72 h. Our study confirms that heparinized blood samples should be stored at RT to get sufficient metaphases for DCA, and that EDTA blood samples also can be used for blood culture after washing and storage under 5.2 ± 1.0 °C.