Claësson M H, Rodger M B, Johnson G R, Whittingham S, Metcalf D
Clin Exp Immunol. 1977 Jun;28(3):526-34.
An improved method is described for growing human T-lymphocyte colonies in agar medium containing phytohaemagglutinin (PHA) and sheep red blood cells (SRBC). Cluster and colony growth was obtained when blood mononuclear cells were plated directly in the agar-medium (one-step procedure) or after incubation of cells in liquid medium with PHA (two-step procedure). In the one-step procedure approximately 1 per 100 cells plated formed a cluster containing four to fifty cells. In the two-step procedure 1 per 20 cells plated formed a cluster or a colony (more than fifty cells). The proliferating cells were shown to be sheep-erythrocyte rosette-forming cells (E-RFC). Optimal proliferation was dependent on the presence of phagocytic cells in the cell suspensions cultured. No growth occurred in cultures depleted of E-RFC. Detailed studies of the cycle, velocity sedimentation, and density of the cells plated showed that the majority of cluster- and colony-forming cells were small non-cycline lymphocytes with a sedimentation velocity of 4 mm/hr, and a density between 1-069 and 1-077 g/cm3.
描述了一种改进的方法，用于在含有植物血凝素（PHA）和绵羊红细胞（SRBC）的琼脂培养基中培养人T淋巴细胞集落。当血液单核细胞直接接种于琼脂培养基中（一步法）或在含PHA的液体培养基中孵育后接种（两步法）时，可获得集簇和集落生长。在一步法中，每接种100个细胞中约有1个形成含有4至50个细胞的集簇。在两步法中，每接种20个细胞中有1个形成集簇或集落（超过50个细胞）。增殖细胞显示为绵羊红细胞花环形成细胞（E-RFC）。最佳增殖依赖于培养的细胞悬液中吞噬细胞的存在。E-RFC耗尽的培养物中无生长。对接种细胞的周期、速度沉降和密度的详细研究表明，大多数集簇和集落形成细胞是小型非循环淋巴细胞，沉降速度为4mm/hr，密度在1.069至1.077g/cm³之间。
