摘要
本文建立了一种改良的适于小鼠脾脏淋巴细胞形成T细胞集落的微量甲基纤维素半固体培养系统。先将脾脏有核细胞(5x10^6/ml)在含有ConA(5μg/ml)的液相中培养24小时,然后收集非粘附细胞,将其接种到含0.88％甲基纤维素、10％FCS,ConA 5μg/ml和15％ConA-LCM的半固体系统中培养,细胞的终浓度为5x10^5/ml。以上培养条件是通过正交设计试验所获得的最优组合,可获得高集落形成率。本文还对集落形成细胞的性质和集落生长的动力学进行了讨论。
A modified methyl cellulose microculture system was developed for T-lymphocyte colony formation from splenic lymphocytes of mice. The cells were preincubated at first in liquid phase for 24 hours at a concentration of 5×106 nucleated cells/ml of RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum (FCS) and concanavalin A (Con A)5μg/ml of culture medium. The non-adhesive cells were then harvested and seeded in semisolid medium containing 0.88% methyl-cellulose in RPMI 1640 supplemented with 10% FCS, Cnn A 10μ g/ml and 15% ConA-stimulated murine lymphocyte conditioned medium at a final cell concentration of 5×105 viable cells/ml. This procedure, as a result of orthogonal tests, was proved to yield highest plating efficiency on 5-7 days of culture.
出处
《杭州大学学报（自然科学版）》
CSCD
1989年第1期77-85,共9页
Journal of Hangzhou University Natural Science Edition
基金
国家自然科学基金资助课题
