Studies on cell interactions using a micromethod for growing T-lymphocyte colonies in agar culture.
Studies on cell interactions using a micromethod for growing T-lymphocyte colonies in agar culture.
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使用微法在琼脂培养物中培养 T 淋巴细胞集落来研究细胞相互作用。
作者：
P. G. Goube de Laforest;P. Thomas;D. Pelletier;J. Tanzer
微法描述了生长pha诱导的t细胞集落从少至5到10×103外周血单核细胞。在标准和微法条件下，不同数量的细胞以相同的浓度在琼脂中镀，其镀效率保持不变。相反，在微培养中获得的细胞剂量响应曲线支持先前的观察结果，即随着细胞浓度达到最佳水平，电镀效率增加。这些曲线的数学分析进一步表明，t细胞集落的形成涉及集落祖细胞（TCFC）和至少两个合作细胞（CC）群体之间的相互作用。原代菌落的细胞仍然含有TCFC，但通常缺乏CC，在传代培养中转移时产生继代菌落，并在培养物中加入CC条件的培养基。在此条件下，随着细胞数的增加，镀效率保持不变，因此，得到的菌落数量与细胞数严格成正比。此外，CC偶尔在原代菌落中持续存在，可以通过(a)在没有CC条件培养基的情况下它们产生继代菌落的能力和(b)随着接种细胞数量的增加而增加的剂量响应曲线的修改来检测。考虑到该系统所需的细胞和条件培养基数量较少，本研究认为该微法为研究t细胞集落形成过程中细胞相互作用提供了一个有用的工具。
A micromethod is described for growing PHA-induced T-cell colonies from as few as 5 to 10×103 peripheral blood mononuclear cells. When various numbers of cells were plated in agar at the same concentration under both standard and micromethod conditions, their plating efficiency remained constant. In contrast, the cell dose—response curves obtained in microculture supported previous observations that plating efficiency increases with cell concentration up to an optimum level. The mathematical analysis of these curves provided further indication that T-cell colony formation involved interactions between a colony progenitor cell (TCFC) and at least two populations of co-operating cells (CC). Pooled cells from primary colonies, which still contained TCFC but were usually depleted of CC, generated secondary colonies when transferred in subculture, provided medium conditioned by CC was added to the cultures. Under these conditions, the plating efficiency remained constant with increasing numbers of cells plated and thus, the number of colonies obtained was found to be strictly proportional to the number of cells plated. In addition, CC persisting occasionally in primary colonies could be detected by (a) their ability to generate secondary colonies in the absence of CC conditioned medium and (b) a modification of the dose—response curves obtained toward increasing plating efficiency with the number of cells seeded. With respect to the small numbers of cells and conditioned media required in this system, it was concluded that the micromethod presented should provide a useful tool for studies on cell interactions during T-cell colony formation.