Maintenance of HSC by Wnt5a secreting AGM-derived stromal cell line
Maintenance of HSC by Wnt5a secreting AGM-derived stromal cell line
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DOI：
10.1016/j.exphem.2010.09.010
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发表时间：
2011-01-01
影响因子：
2.6
通讯作者：
Verfaillie, Catherine M.
中科院分区：
文献类型：
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作者：
Buckley, Shannon M.;Ulloa-Montoya, Fernando;Verfaillie, Catherine M.
目的研究造血干细胞(HSC)维持/扩增HSC的微环境对其分化的影响。方法体外培养UG26-1B6、UG26-1B6、EL08-1D2、胚胎肝细胞系、UG26-1B6细胞系、UG26-1B6细胞系、EL08-1D2细胞系、UG26-1B6细胞系、UG26-1B6细胞系、EL08-1D2细胞系、UG26-1B6细胞系、UG26-1B6细胞系和EL08-1D2细胞系。结果LIN(-)小鼠骨髓细胞在UG26-1B6以上的传代孔中保持3周，而EL08-1D2细胞中含有竞争性再扩增的HSC，在EL08 1D2饲养层中加入25%的UG26-1B6细胞可以维持非接触性培养中的HSC，验证了UG26-1B6细胞分泌可溶性因子，因为我们发现Wnt5a在UG26-1B6细胞中的表达显著高于08 1D2细胞，我们在EL08 1D2 Transwell培养物中加入Wnt5a或在UG26 1B6 Transwell培养物中添加Wnt5a抗体。在EL08-1D2 Transwell培养物中加入Wnt5a恢复了HSC的维持，而在UG26-1B6 Transwell培养物中加入抗Wnt5a抗体则抑制了竞争性再繁殖HSC的维持。结论从胚胎微环境中产生的基质细胞系提供了一种工具来鉴定在维持HSC中起作用的分泌蛋白，并且UG26 1B6细胞产生的至少一个负责维持HSC的因子是Wnt5a(C)2011 ISEH血液学和干细胞学会发表的
Objective The microenvironment wherein hematopoietic stem cells (HSC) reside orchestrates HSC self-renewal vs differentiation decisions Stromal cells derived from ontogenically divergent hematopoietic microenvironments can support HSC in vitro and have been used to decipher factors that Influence HSC fate decisions Employing stromal cell lines derived from the aorta-gonad-mesonephros and embryonic liver, we aim to identify secreted factors that maintain/expand HSC in vitroMaterials and Methods We cultured murine lineage antigen-negative (Lin(-)) bone marrow cells in transwells above the UG26-1B6, urogenital ridge-, and EL08 1D2, embryonic liver-derived cell lines We, also, performed real-time quantitative PCR analysis to identify differentially expressed genes from the Wnt family of proteins in ontogenically different stromal cell linesResults Lin(-) murine bone marrow cells maintained for 3 weeks in transwells above UG26-1B6 but not EL08-1D2 cells contained competitive repopulating HSC Addition of as few as 25% UG26-1B6 cells to EL08 1D2 feeders led to maintenance of HSC in noncontact cultures, validating soluble factors are secreted by the UG26-1B6 cells As we found that Wnt5a was significantly higher expressed in UG26-1B6 than EL08 1D2 cells, we added Wnt5a to EL08 1D2 transwell cultures or an antibody against Wnt5a to UG26 1B6 transwell cultures Addition of Wnt5a to EL08-1D2 transwell cultures restored maintenance of HSC, whereas addition of an anti Wnt5a antibody to UG26-1B6 transwell cultures inhibited maintenance of competitive repopulating HSCConclusions We demonstrate that stromal cell lines generated from embryonic microenvironments provide a tool to identify secreted proteins that play a role in the maintenance of HSC, and that at least one of the factors produced by UG26 1B6 cells responsible for preserving HSC is Wnt5a (C) 2011 ISEH - Society for Hematology and Stem Cells Published by Elsevier Inc