Chen W F, Ewing T, Scollay R, Shortman K
Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia.
Thymus. 1988;12(1):51-68.
A high cloning-efficiency microculture system is described in which single T cells, stimulated to divide by phorbol ester and calcium ionophore, grow rapidly under the influence of purified growth factors in the absence of other cells. The kinetics of clonal growth has been monitored over a five day period by phase-contrast microscopy. Mature peripheral T cells, and mature subpopulations from the thymus, responded with a cloning efficiency over 80%; they required IL-2 as a minimum but several other factors enhanced growth. Ly2+L3T4- thymocytes (mean doubling time 10.4 hr) grew more rapidly than Ly2-L3T4+ thymocytes (mean doubling time 15.2 hr). Early (Ly2-L3T4-) thymocytes responded with a cloning efficiency of 60%; their efficient growth was dependent on both IL-1 and IL-2. The typical Ly2+L3T4+ cortical thymocyte did not grow under these conditions.
本文描述了一种高克隆效率的微培养系统，在该系统中，被佛波酯和钙离子载体刺激而分裂的单个T细胞，在没有其他细胞的情况下，在纯化生长因子的影响下迅速生长。通过相差显微镜在五天时间内监测了克隆生长的动力学。成熟的外周T细胞以及来自胸腺的成熟亚群，克隆效率超过80%；它们至少需要白细胞介素-2，但其他几种因子可促进生长。Ly2 + L3T4 - 胸腺细胞（平均倍增时间10.4小时）比Ly2 - L3T4 + 胸腺细胞（平均倍增时间15.2小时）生长得更快。早期（Ly2 - L3T4 - ）胸腺细胞的克隆效率为60%；它们的高效生长依赖于白细胞介素-1和白细胞介素-2。典型的Ly2 + L3T4 + 皮质胸腺细胞在这些条件下不生长。
