范平生1 朱德发2 陈飞虎3 胡 冰4 林新民4 庄建生4
1安徽省立医院分院肿瘤科
2安徽医科大学附属医院内科
3安徽医科大学药学系
4安徽省立医院肿瘤科
目的：调节细胞培养液pH值，观察不同pH值对LAK 细胞的增殖和rIL-2、LAK细胞、rIL-2 LAK细胞杀伤恶性细胞的影响。方法：采用淋巴细胞分离液梯度离心法从正常人外周血分离单个核细胞并用rIL-2激活，于96孔细胞培养板或培养管中培养。应用细胞计数方法观察pH值分别为6.8、7.3、7.6的培养液，对rIL-2激活单个核细胞的影响。用7404细胞作为靶细胞、MTT法测定培养液pH值分别为6.8、7.3、7.6状态下rIL-2、LAK细胞、rIL-2 LAK细胞的杀伤作用或细胞毒作用。激活后的LAK细胞在不同的pH值条件下与人肝癌细胞7404作用48h后，在倒置显微镜下观察每高倍视野下不典型的E玫瑰花结形成率。 结果：pH值为7.3、7.6状态下，有利于LAK细胞增殖，pH值为7.6状态下rIL-2、LAK细胞、rIL-2 LAK细胞杀伤效果最佳；pH值为6.8状态下几乎无杀伤作用。pH值为6.8、7.6都不利于7404细胞生长，pH值为7.6有利于LAK细胞生长。不换液板中，pH值为7.3时，“LAK细胞 IL-2、 7404细胞”每个高倍视野下，不典型的E玫瑰花结数是pH值为6.8的7倍、是pH值为7.6的2倍。结论：在偏碱性环境下，有利于LAK细胞增殖，并且在此状态下rIL-2、LAK细胞、rIL-2 LAK细胞对肿瘤细胞杀伤效果最佳。pH值为7.3、7.6有利于LAK细胞与靶细胞的接触反应。
FIII 014
Effect of pH Values on LAK Cells Proliferation and Their Anti-tumor Role in Combination with rIL-2
Fan Pingsheng, Zhu Defa, Chen Feihu, Feng Kehai
230031 The tumor internal medicine, Anhui Provincial Ho ital
Objective: By regulating the pH values of the cell culture ,to o erve the change of the proliferation of LAK cells and that of the effectivene of rIL-2 , LAK cells and rIL-2 LAK cells in killing the MTCs. Methods: The single nucleus cells are separated from the normal human peripheral blood through the gradient centrifugation of the lymphocyte separating agent, activated by rIL-2 and cultured in the cell culture dish with 96 holes or the culture tubes to o erve how pH 6.8, 7.3 and 7.6 culture solutio affect the way that rIL-2 activates the single nucleus cells. With cells 7404 as the target cells, the killer role of rIL-2 and cytotoxin of LAK cells in the case of pH 6.8, 7.3 and 7.6 will be a ayed one by one by the MTT method. After the interplay of the activated LAK cells cultured by different pH values with human liver cancer 7404 for 48 hours, the non-typical E rosette forming rate under every high power field of the inverted microscope is o erved. Results: Culture solution 7.3 or 7.6 favors the proliferation of LAK cells, and pH 7.6 yields the optimal lethality on the part of rIL-2, LAK cells and rIL-2 LAK cells while pH 6.8 yields almost no lethality. Neither pH 6.8 nor 7.6 culture solution favors the growth of cells 7404 but pH 7.6 is favorable to the growth of LAK cells. The number of the non-typical E rosette of “LAK cells rIL-2 cells 7404” under every high power field is 7 times as much in the case of pH7.3 as in pH 6.8 and is more than twice as in pH 7.6. Conclusio : pH values of culture solutio affect the proliferation of LAK cells and concerted role of LAK cells and rIL-2 agai t the malignant tumor.. The low-alkali environment favors the LAK cells proliferation and provides the LAK cells and rIL-2 with the optimal state in which to kill the tumor cells. pH 7.3, 7.6 is favorable to the contact reaction of the immunocytes and malignant cells.
FIII 015
原发性骶骨肿瘤及其治疗
房永宏 史宇霞 卫江华 赵志勇 袁勤钊 陈晋丽
03001