ABCG2 expression is correlated neither to side population nor to hematopoietic progenitor function in human umbilical cord blood
ABCG2 expression is correlated neither to side population nor to hematopoietic progenitor function in human umbilical cord blood
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DOI：
10.1016/j.exphem.2008.09.015
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发表时间：
2009-02-01
影响因子：
2.6
通讯作者：
Cross, Michael
中科院分区：
文献类型：
--
作者：
Alt, Rudiger;Wilhelm, Franziska;Cross, Michael
目标。ABCG2转运体已被确定为小鼠骨髓侧群（SP）的决定因素和原始干细胞的潜在标记物。为了评估SP表型和ABCG2表达在人类脐带血（hub）中识别干细胞的潜力，我们直接研究了SP；ABCG2、CD133、CD34的表达；和UCR样本的造血潜能。材料与方法。多色荧光活化细胞分选分析与Hoechst SP程序相结合，允许在新鲜和冷冻保存的UCB细胞中同时检测SP表型以及表面标记。对分选种群进行鹅卵石区域形成细胞（CAFC）活性分析，并通过定量逆转录酶聚合酶链反应检测ABC转运体ABCG2、MDR1和MRP1 mRNA的表达。ABCG2（+）细胞通过磁活化细胞分选富集，以进行严格的分析。hub来源的SP细胞ABCC2阴性，但包含约20%的CD133(+)/CD34（+）细胞。从UCB中分选的SP细胞在第13周的CAFC活性中富集20倍，而磁活化细胞分选富集的ABCG2（+）细胞在CAFC和液体培养中均未保留造血活性。新鲜和低温保存的UCB在SP频率、免疫表型或CAFC电位方面没有显著差异。ABCG2 mRNA在SP中不富集，在CD133细胞中特异性减少了9倍，而MDR1 mRNA在CD133细胞中特异性减少了8倍。我们没有发现ABCG2与hub的SP活性或造血祖细胞功能相关的证据。(C) 2009 ISEH -血液学和干细胞学会。Elsevier Inc.出版。
Objective. The ABCG2 transporter has been identified as a determinant of the side population (SP) in murine bone marrow and a potential marker for primitive stem cells. To assess the potential of the SP phenotype and ABCG2 expression to identify stem cells in human umbilical cord blood (hUCB), we have examined directly the relationship between SP; expression of ABCG2, CD133, and CD34; and hematopoietic potential in UCR samples.Materials and Methods. Multicolor fluoresence activated cell sorting analysis was combined with the Hoechst SP procedure to allow the simultaneous detection of the SP phenotype together with surface markers in cells from fresh and cryopreserved UCB. Sorted populations were analyzed for cobblestone area-forming cell (CAFC) activity and by quantitative reverse transcriptase polymerase chain reaction for expression of mRNA from the ABC transporters ABCG2, MDR1, and MRP1. ABCG2(+) cells were enriched by magnetic-activated cell sorting for stringent analysis.Results. hUCB-derived SP cells were negative for ABCC2, but comprise approximately, 20% CD133(+)/CD34(+) cells. Sorted SP cells from UCB were enriched 20-fold for week 13 CAFC activity, while magnetic-activated cell sorting-enriched ABCG2(+) cells retained no hematopoietic activity either in CAFC or liquid cultures. There was no significant difference in the SP frequency, immunophenotype, or CAFC potential of fresh and cryopreserved UCB. ABCG2 mRNA was not enriched in the SP and was specifically diminished ninefold ill CD133 cells, which were eightfold enriched for MDR1 mRNA.Conclusion. We find no evidence for an association of ABCG2 with SP activity or hematopoietic progenitor function in hUCB. (C) 2009 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc.