Cellular volume and marker expression in human peripheral blood apheresis stem cells
Cellular volume and marker expression in human peripheral blood apheresis stem cells
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DOI：
10.1002/cyto.a.20524
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发表时间：
2008-02-01
期刊：
影响因子：
3.7
通讯作者：
Krishan, Awtar
中科院分区：
文献类型：
--
作者：
Sharma, Siddharth;Cabana, Raquel;Krishan, Awtar
库尔特体积比前向角光散射更准确地测量细胞体积。在本报告中，我们用库尔特体积测定了正常人外周血细胞和血液系统恶性肿瘤患者的造血祖细胞(HPC-A)的平均细胞体积和直径。用CD45-FITC和PE标记的CD34、CD90、CD117和CD133抗体进行CD45-FITC和PE标记的CD34、CD90、CD117和CD133抗体的染色，并进行电子体积和双色荧光分析。新鲜外周血中单个核细胞的平均电子体积和直径分别为淋巴细胞(191µm(3)，7.16µm)、单核细胞(370µm(3)，9.91µm)和粒细胞(328µm(3)，8.56µm)。HIS-3TOPAQUE-1077沉淀法制备的HPC-A单核细胞中，淋巴细胞的体积和直径分别为311微米(3)、8.4微米，单核细胞分别为486微米(3)、9.76微米，粒细胞分别为515微米(3)、9.95微米。10.85亩)。组织不透明-1077沉淀法制备的HPC-A单个核细胞体积与干细胞标志物CD34、CD90、CD117、CD133的表达呈正相关。CD90阳性细胞的平均电子体积最小，为299.93微米(3)，而CD133(322微米(3))、CD117(349微米(3))、CD34(407微米(3))和CD45(453微米(3))阳性细胞的电子体积最小。细胞体积与干细胞标记物表达的相关性可能允许识别小干细胞，而小干细胞可能不表达用于鉴定HPC-A样本中干细胞的常规标记物。(C)2008年国际分析细胞学学会。
Coulter volume is far more accurate measure of cell volume than forward angle light scatter. In this report, we have used Coulter volume to determine the mean cell volume and diameter of normal human peripheral blood cells and hematopoietic progenitor cells obtained by apheresis (HPC-A) from patients with hematological malignancies. Fresh peripheral blood samples (treated with Beckman Coulter IMMUNOPrep erythrocyte lysis solution), HPC-A samples (treated with BD Biosciences FACSLysing solution), or processed by Ficoll Hypaque sedimentation method were stained with CD45-FITC and PE-labeled CD34, CD90, CD117, and CD133 antibodies and analyzed for electronic volume and two color fluorescence. The mean electronic volume and diameter of mononuclear cells from fresh peripheral blood samples prepared with IMMUNOPrep were lymphocytes (191 mu m(3), 7.16 mu m), monocytes (370 mu m(3), 9.91 mu m), and granulocytes (328 mu m(3), 8.56 mu m). In mononuclear cells of HPC-A samples prepared by His-3 topaque-1077 sedimentation, the lymphocytes had volume and diameter of 311 mu m(3), 8.4 mu m, monocytes were 486 mu m(3), 9.76 mu m, and granulocytes were 515 mu m(3), 9.95 mu m. In contrast, HPC-A samples prepared after lysis with FACSLysing solution had mean electronic volume and diameter of lymphocytes (414 mu m(3), 9.25 mu m), monocytes (797 mu m(3), 11.5 mu m), and granulocytes (670 mu m(3), 10.85 mu m). Cell volume of mononuclear cells in the HPC-A samples prepared by Histopaque-1077 sedimentation method was correlated with the expression of stem cell markers CD34, CD90, CD117, and CD133. CD90 positive cells had the smallest mean electronic volume of 299.93 mu m(3)), when compared with cells with positive expression of CD133 (322 mu m(3)), CD 117 (349 mu m(3)), CD34 (407 mu m(3)), and CD45 (453 mu m(3)). Correlation of cell volume with stem cell marker expression may allow for the identification of small stem cells, which may not express the conventional markers used for the identification of stem cells in HPC-A samples. (C) 2008 International Society for Analytical Cytology.