Bártková J, Cinátl J, Novák J T, Mandys V
Folia Biol (Praha). 1982;28(2):124-33.
We pointed out some sources of variations in the results obtained by the method of human peripheral lymphocyte culture in semi-solid medium. The influence of changes in five important parameters of the two-layer method was studied: agar concentration in the upper layer, cell concentration, mitogen dilution, type of culture medium and serum. We found that the optimal conditions for lymphocyte culture and for the evaluation of the results were 0.3% agar concentration in the upper layer, the concentration of 5 x 10(5) cells/ml, and a double dilution of phytohaemagglutinin. Of the culture media tested, MEM with 0.2% Bactotryptose supplemented with 20% inactivated mixed human serum proved most appropriate. Cells in lymphocyte colonies were characterized morphologically, cytochemically and by means of basic membrane features. The possibilities of using the method in clinical work are shown and the necessity of standardizing the culture technique is emphasized.
我们指出了在半固体培养基中采用人外周血淋巴细胞培养法所获结果的一些变异来源。研究了双层法五个重要参数变化的影响：上层琼脂浓度、细胞浓度、促有丝分裂原稀释度、培养基类型和血清。我们发现淋巴细胞培养及结果评估的最佳条件为上层琼脂浓度0.3%、细胞浓度5×10⁵个/ml以及植物血凝素双倍稀释。在所测试的培养基中，添加20%灭活混合人血清的含0.2%细菌胰蛋白胨的MEM培养基最为合适。对淋巴细胞集落中的细胞进行了形态学、细胞化学及基本膜特征方面的鉴定。展示了该方法在临床工作中的应用可能性，并强调了规范培养技术的必要性。
