Enrichment of human hematopoietic stem cell activity in the CD34+Thy-1+Lin- subpopulation from mobilized peripheral blood.
Enrichment of human hematopoietic stem cell activity in the CD34+Thy-1+Lin- subpopulation from mobilized peripheral blood.
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DOI：
10.1182/blood.v85.2.368.bloodjournal852368
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发表时间：
1995-01
期刊：
影响因子：
20.3
通讯作者：
Lesley J. Murray;B. Chen;A. Galy;S. Chen;R. Tushinski;N. Uchida;R. Negrin;G. Tricot;S. Jagannath;D. Vesole
中科院分区：
文献类型：
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作者：
Lesley J. Murray;B. Chen;A. Galy;S. Chen;R. Tushinski;N. Uchida;R. Negrin;G. Tricot;S. Jagannath;D. Vesole
已知癌症患者的外周血中的CD34+细胞的数量在施用高剂量化疗和造血生长因子后增加。这些所谓的外周血干细胞移植物现在经常用于恶性肿瘤患者的自体移植。在这份报告中，我们解决的问题，是否真正的长期再生多能造血干细胞（PHSC）动员到外周血化疗加粒细胞/巨噬细胞集落刺激因子（GM-CSF）或粒细胞集落刺激因子（G-CSF）动员。我们通过使用抗人Thy-1分子的抗体对CD34+谱系-（Lin-）群体染色，检查了动员的外周血（MPB）中干细胞的存在。研究了动员CD34 + Thy-1 + Lin-细胞进入外周血的动力学，发现具有这种表型的细胞的百分比根据白细胞去除术的天数而变化很大。通过荧光激活细胞分选（FACS）分离可能含有PHSC的CD34 + Thy-1 + Lin-细胞群，并分析其活性。使用体外长期培养系统确定MPB中该候选含干细胞群体的多谱系分化能力，其中鹅卵石区域形成用作检测PHSC的手段。我们还测量了通过使用两种体内模型，其中严重联合免疫缺陷（SCID）-hu小鼠植入人胎骨或胸腺移植物的重建能力。使用这些试验，我们表明，最高频率的鹅卵石区域形成细胞（CAFC）培养7周后观察到的亚群的CD34 + Lin-细胞，表达低水平的Thy-1。该细胞群能够产生B和髓样细胞，并在这些长期培养物中维持CD 34 + Lin-细胞。此外，CD34 + Thy-1 + Lin-细胞亚群具有更高的植入能力，并在SCID-hu骨测定中在8周时显示出多系分化潜能。然而，在SCID-hu胸腺模型中，Thy-1+和Thy-1-亚群均能够在6周时植入供体T细胞，表明两个群体中均存在能够启动T淋巴细胞生成的细胞。（400字处删节）
The number of CD34+ cells in the peripheral blood of cancer patients is known to be increased following the administration of high dose chemotherapy and hematopoietic growth factors. These so-called peripheral blood stem cell grafts are now frequently used for autologous transplantation of patients with malignancies. In this report, we address the question of whether true long-term repopulating pluripotent hematopoietic stem cells (PHSC) are mobilized into peripheral blood following chemotherapy plus granulocyte/macrophage colony-stimulating factor (GM-CSF) or granulocyte colony-stimulating factor (G-CSF) mobilization. We have examined the presence of stem cells in mobilized peripheral blood (MPB) by using an antibody to the human Thy-1 molecule to stain the CD34+Lineage- (Lin-) population. The kinetics of mobilization of CD34+Thy-1+ Lin- cells into peripheral blood were studied, and the percentage of cells with this phenotype was found to vary widely depending on the day of leukapheresis. A CD34+Thy-1+Lin- cell population, potentially containing PHSCs, was isolated by fluorescence activated cell sorting (FACS) and analyzed for activity. The multilineage differentiative capacity of this candidate stem cell-containing population in MPB was determined using an in vitro long-term culture system, in which cobblestone area formation was used as a means of detecting PHSCs. We also measured repopulating capacity by using two in vivo models in which severe combined immunodeficiency (SCID)-hu mice were implanted with human fetal bone or thymus grafts. Using these assays, we show that the highest frequency of cobblestone area-forming cells (CAFC) after 7 weeks of culture was observed in a subpopulation of CD34+Lin- cells, which expressed low levels of Thy-1. This cell population was capable of producing both B and myeloid cells, and maintaining CD34+Lin- cells in these long term cultures. Moreover, the CD34+Thy-1+Lin- cell subset possessed a higher ability to engraft and to demonstrate multilineage differentiative potential at 8 weeks in the SCID-hu bone assay. However, in the SCID-hu thymus model, both Thy-1+ and Thy-1- subpopulations were capable of donor T-cell engraftment at 6 weeks, suggesting the presence of cells capable of initiating T lymphopoiesis in both populations.(ABSTRACT TRUNCATED AT 400 WORDS)