【摘要】 本文报道了应用国产2-氨乙基异硫脲溴化物(AET)处置绵羊红细胞形成玫瑰花结,用以提纯T淋巴细胞约在95%左右,提高了国内所分离T淋巴细胞的纯度。并讨论了其可能的原因。更多还原

【Abstract】 Periferal bloodfrom 15 healthy blood donors were purified for isolating lymphocytes, After the application of initial centrifugation of heparinized venous blood over a ficoll-isopague (FIP) gradient the interphase cells were collected and washed, Depletion of monocytes was carried out by means of adherence to plastic petri dishes in which the cells were cultured.The separation of T and B cell enriched fractions was carried out by rosetting with AET treated SRBC followed by gradient centrifugation procedure. Lymphocytes at a concentration of 10～15×10~6 cells/ml in RPM1 medium were mixed with equal volumes of 1% AET treated SRBC in 100% heat-inactivated SRBC-absorbed FCS, After centrifugation and incubation in 37℃ for 5 minutes the tubes were placed at 4℃ over night Mean while E rosettes with both SRBC and AET treated SRBC were manipulated,It was shown that the E-rosette percentages of AET-SRBC rosette with lymphocytes were significantly higher than that of non-AET group. It was also shown that approximatly 95% of T lymphocytes were purified by the method used.更多还原