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本文报道用PHA作为刺激物,采用简化的一步法在甲基纤维素培养基上培养人外周血T淋巴细胞集落(TL一CFU)获得成功,并对TL一CFU培养的适宜条件进行了探讨;同时,对集落组成细胞进行了T淋巴细胞表型分析;在此适宜条件下,对健康成人外周血TL—CFU产率进行了测定。结果显示:(1)培养体系中PHA2ul/ml、20%小牛血清(FCS)、5×10(-5)M2一巯基乙醇、0.8%甲基纤维素、60%RPMI1640、2×105MNC/ml、培养时间七天为TL—CFU培养的适宜条件;(2)集落组成细胞中CD3+细胞占92.5±4.59%、CD4+、CD8+细胞分别为57.83±7.85%及46.66±4.76%、CD4/CD8之比为1.24±0.17;(3)健康成人外周血TL一CFU产率为351.58±21.97/2×105MN。基于上述结果认为,该培养方法操作简便、集落产率高,为研究T淋巴细胞及其亚群提供了条件。
This article reports the success of culturing human peripheral blood T lymphocyte colonies (TL-CFU) on a methylcellulose medium by using a simplified one-step method with PHA as a stimulator, and discusses suitable conditions for culturing the TL-CFU. At the same time, T lymphocyte phenotype analysis of colony-forming cells was carried out. Under this suitable condition, the yield of TL-CFU in peripheral blood of healthy adults was measured. The results showed that: (1) PHA 2ul / ml, 20% FCS, 5 × 10 (-5) M mercaptoethanol, 0.8% methylcellulose, 60% RPMI1640, 2 × 105MNC / Ml, culture time seven days for the TL-CFU culture conditions; (2) CD3 + cells accounted for 92.5 ± 4.59%, CD4 +, CD8 + cells were 57.83 ± 7.85% and 46 .66 ± 4.76%, the ratio of CD4 / CD8 was 1.24 ± 0.17; (3) The yield of TL-CFU in peripheral blood of healthy adults was 351.58 ± 21.97 / 2 × 105MN. Based on the above results, this method is simple and easy to operate, with high yield of colonies, which provides conditions for the study of T lymphocytes and their subpopulations.