Jiang Mengmeng, Chen Haide, Lai Shujing, Wang Renying, Qiu Yunfei, Ye Fang, Fei Lijiang, Sun Huiyu, Xu Yang, Jiang Xinyi, Zhou Ziming, Zhang Tingyue, Li Yanwei, Xie Jin, Fang Qun, Gale Robert Peter, Han Xiaoping, Huang He, Guo Guoji
1Center for Stem Cell and Regenerative Medicine, Zhejiang University School of Medicine, Zhejiang, Hangzhou, 310058 China.
2Stem Cell Institute, Zhejiang University, Zhejiang, Hangzhou, 310058 China.
Cell Discov. 2018 Oct 30;4:59. doi: 10.1038/s41421-018-0059-5. eCollection 2018.
Identification of effective culture conditions to maintain and possibly expand human HSPCs in vitro is an important goal. Recent advances highlight the efficacy of chemicals in maintaining and converting cell fates. We screened 186 chemicals and found that a combination of CHIR-99021, Forskolin and OAC1 (CFO) maintained human CD34-positive cells in vitro. Efficiency of the culture system was characterized using flow cytometry for CD34-positive cells, a colony-forming assay and xeno-transplants. We found that human CD34-positive cells treated with this combination had enhanced expression of human HSPC markers and increased haematopoietic re-populating ability in immune-deficient mice. Single-cell RNA-seq analyses showed that the in vitro cultured human CD34-positive cells were heterogeneous. We found that CFO supports maintenance of human CD34-positive cells by activating , and AKT-cAMP signaling pathway. These data have implications in therapies requiring maintenance and/or expansion of human HSPCs.
确定有效的培养条件以在体外维持并可能扩增人造血干细胞（HSPCs）是一个重要目标。最近的进展突出了化学物质在维持和转变细胞命运方面的功效。我们筛选了186种化学物质，发现CHIR-99021、福斯高林和OAC1（CFO）的组合可在体外维持人CD34阳性细胞。使用针对CD34阳性细胞的流式细胞术、集落形成试验和异种移植对培养系统的效率进行了表征。我们发现，用这种组合处理的人CD34阳性细胞在免疫缺陷小鼠中增强了人HSPC标志物的表达并提高了造血重建能力。单细胞RNA测序分析表明，体外培养的人CD34阳性细胞是异质性的。我们发现CFO通过激活 、 和AKT-cAMP信号通路来支持人CD34阳性细胞的维持。这些数据对需要维持和/或扩增人HSPCs的治疗具有启示意义。
Zotero 插件
