Abstract

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 HOXA9, GATA2 and AKT-cAMP signaling pathway. These data have implications in therapies requiring maintenance and/or expansion of human HSPCs.

Details

Title
Maintenance of human haematopoietic stem and progenitor cells in vitro using a chemical cocktail
Author
Jiang, Mengmeng 1 ; Chen, Haide 1 ; Lai, Shujing 1 ; Wang, Renying 1 ; Qiu, Yunfei 2 ; Ye, Fang 1 ; Lijiang Fei 1 ; Sun, Huiyu 1 ; Xu, Yang 3 ; Jiang, Xinyi 1 ; Zhou, Ziming 1 ; Zhang, Tingyue 1 ; Li, Yanwei 4 ; Xie, Jin 5 ; Fang, Qun 6 ; Gale, Robert Peter 7 ; Han, Xiaoping 1 ; Huang, He 8 ; Guo, Guoji 9 

 Center for Stem Cell and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou, China; Stem Cell Institute, Zhejiang University, Hangzhou, China 
 Institute of Hematology, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China 
 UT-ORNL Graduate School of Genome Science and Technology, The University of Tennessee, Knoxville, TN, USA 
 Core Facilities, Zhejiang University School of Medicine, Hangzhou, China 
 Institute of Mechatronic Control Engineering, Zhejiang University, Hangzhou, China 
 Department of Chemistry, Institute of Microanalytical Systems, Zhejiang University, Hangzhou, China 
 Department of Medicine, Haematology Research Centre, Division of Experimental Medicine, Imperial College London, London, UK 
 Stem Cell Institute, Zhejiang University, Hangzhou, China; Institute of Hematology, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China; Alliance for Atlas of Blood Cells, Hangzhou, China 
 Center for Stem Cell and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou, China; Stem Cell Institute, Zhejiang University, Hangzhou, China; Institute of Hematology, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China; Alliance for Atlas of Blood Cells, Hangzhou, China; Zhejiang Provincial Key Lab for Tissue Engineering and Regenerative Medicine, Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cell and Regenerative Medicine, Hangzhou, China 
Pages
1-12
Publication year
2018
Publication date
Oct 2018
Publisher
Springer Nature B.V.
e-ISSN
20565968
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2126882937
Copyright
© 2018. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.