Endothelial Cells Promote Expansion of Long-Term Engrafting Marrow Hematopoietic Stem and Progenitor Cells in Primates.
Endothelial Cells Promote Expansion of Long-Term Engrafting Marrow Hematopoietic Stem and Progenitor Cells in Primates.
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DOI：
10.5966/sctm.2016-0240
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发表时间：
2017-03
影响因子：
6
通讯作者：
Kiem HP
中科院分区：
文献类型：
--
作者：
Gori JL;Butler JM;Kunar B;Poulos MG;Ginsberg M;Nolan DJ;Norgaard ZK;Adair JE;Rafii S;Kiem HP
骨髓（BM）造血干细胞和祖细胞（HSPC）的成功扩增将有益于许多HSPC移植和基因治疗/编辑应用。然而，目前的扩增技术受到离体多能性和自我更新特性丧失的限制。我们假设离体血管生态位将提供促造血信号以扩增HSPC，同时保持多能性和自我更新。为了验证这一假设，将BM自体CD 34+细胞在内皮细胞（EC）共培养物中扩增并移植到非人灵长类动物中。与EC共培养的CD 34 +C38− HSPC扩增高达17倍，与单独使用细胞因子培养的细胞相比，造血集落形成活性显著增加（集落形成单位-粒细胞-红细胞-巨噬细胞-单核细胞; p < .005）。用绿色荧光蛋白慢病毒载体转导并在EC上扩增的BM CD 34+细胞，用多谱系多克隆重建长期移植。在粒细胞、淋巴细胞、血小板和红细胞中观察到基因标记。全转录组分析表明，EC共培养改变了BM CD 34+细胞中75个基因的表达谱，而不妨碍长期植入潜力。这些发现表明，离体血管生态位是扩增成年BM HSPC的有效平台。干细胞转化医学2017;6：864-876
Successful expansion of bone marrow (BM) hematopoietic stem and progenitor cells (HSPCs) would benefit many HSPC transplantation and gene therapy/editing applications. However, current expansion technologies have been limited by a loss of multipotency and self‐renewal properties ex vivo. We hypothesized that an ex vivo vascular niche would provide prohematopoietic signals to expand HSPCs while maintaining multipotency and self‐renewal. To test this hypothesis, BM autologous CD34+ cells were expanded in endothelial cell (EC) coculture and transplanted in nonhuman primates. CD34+C38− HSPCs cocultured with ECs expanded up to 17‐fold, with a significant increase in hematopoietic colony‐forming activity compared with cells cultured with cytokines alone (colony‐forming unit‐granulocyte‐erythroid‐macrophage‐monocyte; p < .005). BM CD34+ cells that were transduced with green fluorescent protein lentivirus vector and expanded on ECs engrafted long term with multilineage polyclonal reconstitution. Gene marking was observed in granulocytes, lymphocytes, platelets, and erythrocytes. Whole transcriptome analysis indicated that EC coculture altered the expression profile of 75 genes in the BM CD34+ cells without impeding the long‐term engraftment potential. These findings show that an ex vivo vascular niche is an effective platform for expansion of adult BM HSPCs. Stem Cells Translational Medicine 2017;6:864–876