Sych F J
Allerg Immunol (Leipz). 1981;27(2):102-23.
A simple routine method for the phytomitogen-(PHA-, Con A-) induced stimulation of purified rabbit peripheral blood lymphocytes is described. The lymphocytes are cultivated in closed polypropylene Eppendorf tubes at 37 degrees C, the mitogen answer is quantified by determination of 3H-thymidine (3HT) incorporation by means of liquid scintillation spectrometry of cell-sediment layered filter paper discs. The optimal experimental conditions for each step of the test procedure are given. The mean variation coefficients of triplicate determinations of non-stimulated negative controls (n = 28) and mitogen-activated lymphocyte cultures (n = 56) are 17.0 (6-28) and 5.6 (1-13)%, respectively.
本文描述了一种用于植物有丝分裂原(PHA、Con A)诱导纯化兔外周血淋巴细胞刺激的简单常规方法。淋巴细胞在封闭的聚丙烯Eppendorf管中于37℃培养,通过对细胞沉淀分层滤纸圆盘进行液体闪烁光谱法测定3H-胸腺嘧啶核苷(3HT)掺入量来定量有丝分裂原反应。给出了测试程序各步骤的最佳实验条件。未刺激的阴性对照(n = 28)和有丝分裂原激活的淋巴细胞培养物(n = 56)的一式三份测定的平均变异系数分别为17.0(6 - 28)%和5.6(1 - 13)%。