流式细胞术联合检测骨髓细胞CD271、CD133、CD34的表达与分析

本研究检测骨髓细胞的CD271、CD133和CD34的表达，分析细胞表达CD271与CD133及CD133与CD34的相关性。根据不同设计组合用CIN5-PerCP、CD271-PITC、CD133-PE和CD34-FITC标记骨髓细胞，获得细胞后以FSC、SSC和CD45反复对细胞群进行定位和筛选，然后用流式细胞术检测骨髓细胞和骨髓单个核细胞（MNC）的上述表达。结果表明，骨髓细胞CD271^＋、CD133^＋和CD34^＋表达率分别为0．16％、0．20％和0．43％，骨髓单个核细胞分离后CD271^＋、CD133^＋和CD34^＋表达率分别0．49％、0．47％和1．07％；骨髓细胞的CD271^＋CD133^共表达率为0．02％，单个核细胞分离后的CD271^＋CD133^＋的共表达率为0．03％。90％CD133^＋细胞表达CD34，40％CD34^＋细胞表达CD133。结论：建立的三色荧光联合检测方法可作为检测骨髓中CD271^＋，CD133^＋和CD34^＋细胞的方法。CD271阳性细胞与CD133和／或CD34阳性细胞是不同的细胞群，CD271可能在评价和指导骨髓间充质干细胞的临床治疗中有重要意义。

Detection and Analysis of CD271,CD133 and CD34 Expressions in Bone Marrow Cells by Flow Cytometry with Three Color Fluorescence Labelling

This study was purposed to detect the expressions of CD271, CD133 and CD34, and to analyze the correlation of CD271 with CD133 and CD133 with CD34 expressions. The human bone marrow cells （BMCs） and mononucleated cells （MNCs） were detected by flow cytomotry with CD45-PerCP, CD271-FITC, CD133-PE and CD34-FITC labelling according to different combinations of design, cells were located and selected repeatly by FSC, SSC and CD45 after acquirement, then the expressions of CD271, CD133 and CD34 were detected by flow cytometry. The results showed that the expressions of CD271, CD133 and CD34 in BMCs were 0. 16%, 0.20% and 0.43% respectively, while their expressions were 0.49%, 0.47% and 1.07% respectively after isolation of MNCs. The co-expressions of CD271^＋ CD133^＋ before and after isolation of MNCs were （0.02±0.01 ） % and （0.03±0.02） % respectively. The coexpression of CD133^＋ and CD34^＋ before and after isolation of MNCs were （0.18±0.11 ） % and （0.42±0.23 ） % respectively （p 〈0.01 ） ; meanwhile about 90% of cells with CD133^＋ expressed CD34 and 40% of cells with CD34^＋ expressed CD133. It is concluded that the established method of detection using flow cytemetry with three color fluorescence labelling can be used to detect expression of CD271, CD133 and CD34 in BMCs. The cells with CD271 are different from cells with CD133 and CD34, which suggests that the CD271 may be of important role in evaluating and guid- ing the clinical application of BM MSCs.