CD34-positive cells and their subpopulations characterized by flow cytometry analyses on the bone marrow of healthy allogenic donors
CD34-positive cells and their subpopulations characterized by flow cytometry analyses on the bone marrow of healthy allogenic donors
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DOI：
10.1590/s1516-31802009000100004
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发表时间：
2009-01-01
影响因子：
1.4
通讯作者：
Oliveira, José Salvador Rodrigues de
中科院分区：
文献类型：
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作者：
Carvalho, Jerusa Martins;Souza, Marlon Knabben de;Oliveira, José Salvador Rodrigues de
背景与目的：计数和分离不同来源的造血干细胞对于研究和临床检测具有重要意义。我们的目的是描述和量化骨髓供体中的造血细胞群，评估CD 34表达并将其与植活相关联。设计和设置：对圣保罗两家医院的同种异体移植患者进行造血干细胞测定的横断面研究，采用流式细胞术对供体骨髓样本进行分析。根据CD34FTIC、CD117PE、CD38PE、CD7FITC、CD33PE、CD10FITC、CD19PE、CD14FITC、CD13PE、CD11cPE、CD15FITIC、CD22PE、CD 61 FITC和CD 56 PE单克隆抗体在CD 45 PerCP+细胞中的表达，寻找分化和成熟区域。分析⑶ 34+分选细胞的⑶ 38和⑶ 117。在分选之前和之后进行Rh-123保留。结果：R1区CD 34 +/CD 45+细胞占0.1%~ 2.8%，CD 34 +/CD 45-细胞占1.1%; R2中CD 45+细胞主要共表达CD 38、CD 22、CD 19和CD 56，R3和R4中主要共表达CD 33、CD 11 c、CD 14、CD 15和CD 61。分选后，2.2x10（6）个CC 34+细胞相当于总细胞的4.9%。CD 34 +/CD 38+和CD 34 +/CC 117+共表达分别发生在94.9%和82%的事件中。CD 34+细胞与移植物植入呈正相关。超过80%的骨髓细胞高表达Rh-123。CD 34+细胞分选结果显示，分化程度较高的细胞比原始细胞更能保留Rh-123.结论：真正的干细胞是CD 34 +/CD 45-/CD 38-/low-Rh-123的聚集.
CONTEXT AND OBJECTIVE: Counting and separating hematopoietic stem cells from different sources has importance for research and clinical assays. Out aims here were to characterize and quantify hematopoietic cell populations in marrow donors and to evaluate CD34 expression and relate this to engraftment.DESIGN AND SETTING: Cross-sectional study on hematopoietic stem cell assays, using flow cytometry on donor bone marrow samples, for allogenic transplantation patients at two hospitals in Sao Paulo.METHODS: Immunophenotyping of marrow cells was performed in accordance with positive findings of CD34FTIC, CD117PE, CD38PE, CD7FITC, CD33PE, CD10FITC, CD19PE, CD14FITC, CD13PE, CD11cPE, CD15FITIC, CD22PE, CD61FITC and CD56PE monoclonal antibodies in CD45PerCP+ cells, searching for differentiation and maturation regions. CD34+ sorting cells were analyzed for CD38 and CD117. Rh-123 retention was done before and after sorting. Antigen expression and CD34+ cells were correlated with engraftment.RESULTS: In region R1, 0.1% to 2.8% of cells were CD34+/CD45+ and 1.1%, CD34+/CD45-. The main coexpressions of CD45+ cells were CD38, CD22, CD19 and CD56 in R2 and CD33, CD11c, CD14, CD15 and CD61 in R3 and R4. After sorting, 2.2x10(6) CC34+ cells were equivalent to 4.9% of total cells. Coexpression of CD34+/CD38+ and CD34+/CC117+ occurred in 94.9% and 82% of events, respectively. There was a positive relationship between CD34+ cells and engraftment. More than 80% of marrow cells expressed high Rh-123. CD34+ cell sorting showed that cells in regions of more differentiated lineages retained Rh-123 more intensively than in primitive lineage regions.CONCLUSION: We advocate that true stem cells are CD34+/CD45-/CD38-/low-Rh-123 accumulations.