【作者】 赵黎； 孙红英； 张洁； 王夏夏； 刘瑾；

【Author】 ZhaoLi;Sun Hongying;Zhang jie;Wang Xiaxia;Liu Jin;Huashan Hospital, Fudan University;

【机构】 复旦大学附属华山医院；

【摘要】 目的:探讨OLP病人外周血T淋巴细胞分选、培养及纯化技术,探究T淋巴细胞体外生长周期、增殖及方凋亡状态,为细胞共培养提供条件。方法:采用密度梯度离心和免疫磁珠特异性分选,获得T淋巴细胞,检测其分选后T淋巴细胞纯度,并监测T淋巴细胞体外培养中亚群变化和凋亡状态,并绘制生长曲线。结果:T细胞分选纯度可达97.7%。OLPT淋巴细胞的生长曲线基本呈"S"型,传代周期约4-6d,体外培养可至第3—4代。随着体外培养时间增加细胞活力减低,细胞凋亡增加。结论:从OLP外周血中分离出高纯度及高活力的T淋巴细胞,并可成功进行体外培养20天,为后续细胞共培养提供了实验依据。更多还原

【Abstract】 Objective:Study the cell sorting, culture and purification of T lymphocytes in peripheral blood of OLP patients, and to explore the growth cycle, proliferation and apoptosis of T lymphocytes in vitro, providing information for cell co-culture. Methods:Using Density gradient centrifugation and immunomagnetic sorting to obtain T lymphocyte and tested the purity. We detected the change of T lymphocyte subgroup and apoptosis condition andthen drew the growth curve. Results: The purity of Tlymphocyte was up to 97.7%.Cell growth curve of T lymphocyte from OLP patients was ‘S’ shape, cell passage cycle was 4-6 d and the T lymphocyte could be maintained up to 3-4 generations in vitro. Cell viability decreased and apoptosis increased with increasing culture timein vitro. Conclusion : We had successfully obtained T lymphocyte with high purity and viability from peripheral bold of OLP patients, and cultured successfully for 20 daysin vitro, which provided experimental basis for subsequent cell co-culture.更多还原