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林壽德,全茂炯,尹在一,韓乙男,夏潤文 WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 1981 東西醫學硏究所 論文集 Vol.1 No.1
A human fibroblast cell strain, GM 2767, selected to use for human interferon bioassay was investigated on the in vitro biological properties; morphology, growth cycle under various culture conditions, cryopreservation and sensitivity to vesicular stomatitis virus. The results obtained are summarized as follows; 1. The uninactivated fetal bovine serum(FBS) supplemented in the medium resulted in higher growth rate of GM 2767 than the inactivated FBS. The higher concentration of the uninactivated FBS in the medium induced the more stimulated growth rate. Cell growth rate was lower in pH 6.8 and 7.8 than in pH 7.3 of medium. 2. Growth rate was gradually decreased as the passage level was increased. It took more than 7 days to form full monolayer at the passage levles after the 11th. 3. In comparison of two cryopreservative agents of glycerol and dimethyl sulfoxide(DMSO), no difference was found in preservation of cell viability. However, the plating efficiency of the preserved cells was higher in DMSO than in glycerol. 4. GM 2767 cells was highly susceptible to vesicular stomatitis virus(VSV) challenge, showing typical cytopathic effects of VSV at 12-36 hours postinoculation.
免疫生物學的 硏究에 應用되는 Cell Line들의 保存 關한 硏究 Ⅱ. Natural Killer Cell Assay 標的 細胞로 利用되는 K 562 細胞 및 培養 및 牲狀
林壽德,金光爀,尹在一,金光爀,韓乙男,夏潤文 WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 1981 東西醫學硏究所 論文集 Vol.1 No.1
Natural killer 細胞檢定에서 標的 細胞로 利用하기 위해서 慢性骨髓性白血病患者에서 얻어진 K562 cell line 을 日本에서 導入하였으며 이 細胞에 대한 培養條件, 形態學的 性狀 및 長期冷凍保存에 대해 연구하였다. 또한 本 硏究所에서 適應되고 數代 繼代培養된 K562 細胞의 正常人 NK 細胞에 대한 感受性을 試驗하였으며 K562 細胞의 生物學的 性狀에 대해 考察하였다. 實驗을 통해 얻어진 結果를 要約하면 다음과 같다. 1. K562 細胞는 多樣形態性을 나타냈다. 培養初期에는 均一한 形態를 보였으나 時間이 경과함에 따라 多樣하게 變化하였다. 細胞의 크기는 正常人 淋巴球의 1.5培 내지 3.8培에 달했으며 核分裂染色體가 현저히 많이 관찰 됐다. 細胞는 均一한 浮遊狀態로 增殖했다. 2. K562 細胞의 增殖率은 RPMI 1640 및 MEM 培地에서보다 enriched RPMI 1640 에서 높게 나타났다. 또한 10% 내지 15% 牛胎兒血淸濃度와 5% CO₂-95% air 狀態下에서 增殖率이 높았다. 3. 細胞增殖率은 培養開始 때 細胞濃度가 ?? 일때 높았고 細胞의 viability 가 높을수록 增殖能도 높았다. 4. 液體窒素內 冷凍保存時 K562 細胞의 viability 는 약 29.2% 減少하였다. 5. Trypan blue exclusion 方法에 의해 K562 細胞의 正常人 NK 細胞에 대한 感受性을 試驗한 바, effector/target 비율이 5:1 일때 1:1 및 2:1 비율에서 보다 높아서 男子는 33.3%, 女子는 30.5%로 나타났다.
韓國正常人의 末稍血液淋巴球의 Natural Kiier(NK)活性値에 關한 硏究
夏潤文,金光爀,全茂炯,禹種設,林壽德 WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 1981 東西醫學硏究所 論文集 Vol.1 No.1
Spontaneous human lymphocyte-mediated cytotoxicity(SLMC) against target of cultured myeloid cell(K562) was examined in healthy subjects of Korean. Lymphocyte-mediated natural cytotoxicity against target cell was assayed by using a ?? release microcytotoxicity system. These assay system involve reactivity in 37℃, CO₂ contained incubator between effector cell(PBL) and target cell, K562 at 40 to 1 of L/T ratio. The SLMC of lymphocytes from healthy donors were compared between the male and female. The result showed differences of certain degree in activity in both group. In our study, the results of NK activity in male and female are definitely different. This results was consistent with other investigators.