Mesenchymal Stem Cells Secreting Angiopoietin-Like-5 Support Efficient Expansion of Human Hematopoietic Stem Cells Without Compromising Their Repopulating Potential
Mesenchymal Stem Cells Secreting Angiopoietin-Like-5 Support Efficient Expansion of Human Hematopoietic Stem Cells Without Compromising Their Repopulating Potential
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DOI：
10.1089/scd.2010.0456
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发表时间：
2011-08-01
影响因子：
4
通讯作者：
Chen, Jianzhu
中科院分区：
文献类型：
--
作者：
Khoury, Maroun;Drake, Adam;Chen, Jianzhu
人类造血干细胞（HSC）的临床和临床前应用常常受到细胞稀缺的限制。因此，扩大人类造血干细胞以增加其数量，同时保持其干细胞特性已成为一个重要的研究领域。在此，我们报告了一种强大的HSC共培养系统，其中脐带血CD34（+）CD133（+）细胞与经工程改造以在确定的培养基中表达血管生成素样5的间充质干细胞共培养。培养 11 天后，通过有限稀释测定，在 NOD-scid Il2rg(-/-) (NSG) 小鼠中，SCID 再生细胞扩增了约 60 倍。在竞争性再增殖实验中，培养的CD34+CD133+细胞与未培养的CD34+CD133+细胞具有相似的植入潜力，并且能够进行有效的二次重建。此外，扩增的细胞支持 NSG 受体小鼠中人血细胞的稳健多谱系重建，包括更有效的 T 细胞重建。这些结果表明，扩增的CD34(+) CD133(+)细胞维持短期和长期HSC活性。据我们所知，这种类似于 SCID 再生细胞 60 倍的扩增是迄今为止报道的人类 HSC 的最佳扩增。因此，有必要进一步开发这种共培养方法，以扩大人类造血干细胞的临床和临床前应用。
Clinical and preclinical applications of human hematopoietic stem cells (HSCs) are often limited by scarcity of cells. Expanding human HSCs to increase their numbers while maintaining their stem cell properties has therefore become an important area of research. Here, we report a robust HSC coculture system wherein cord blood CD34(+) CD133(+) cells were cocultured with mesenchymal stem cells engineered to express angiopoietin-like-5 in a defined medium. After 11 days of culture, SCID repopulating cells were expanded similar to 60-fold by limiting dilution assay in NOD-scid Il2rg(-/-) (NSG) mice. The cultured CD34(+) CD133(+) cells had similar engraftment potential to uncultured CD34(+) CD133(+) cells in competitive repopulation assays and were capable of efficient secondary reconstitution. Further, the expanded cells supported a robust multilineage reconstitution of human blood cells in NSG recipient mice, including a more efficient T-cell reconstitution. These results demonstrate that the expanded CD34(+) CD133(+) cells maintain both short-term and long-term HSC activities. To our knowledge, this similar to 60-fold expansion of SCID repopulating cells is the best expansion of human HSCs reported to date. Further development of this coculture method for expanding human HSCs for clinical and preclinical applications is therefore warranted.