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Hypoxia with Wharton’s jelly mesenchymal stem cell coculture maintains stemness of umbilical cord blood-derived CD34(+) cells

BACKGROUND: The physiological approach suggests that an environment associating mesenchymal stromal cells with low O(2) concentration would be most favorable for the maintenance of hematopoietic stem/progenitor cells (HSPCs). To test this hypothesis, we performed a coculture of cord blood CD34(+) ce...

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Autores principales: Zhao, Dewan, Liu, Lingjia, Chen, Qiang, Wang, Fangfang, Li, Qiuyang, Zeng, Qiang, Huang, Jingcao, Luo, Maowen, Li, Wenxian, Zheng, Yuhuan, Liu, Ting
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5998541/
https://www.ncbi.nlm.nih.gov/pubmed/29895317
http://dx.doi.org/10.1186/s13287-018-0902-5
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author Zhao, Dewan
Liu, Lingjia
Chen, Qiang
Wang, Fangfang
Li, Qiuyang
Zeng, Qiang
Huang, Jingcao
Luo, Maowen
Li, Wenxian
Zheng, Yuhuan
Liu, Ting
author_facet Zhao, Dewan
Liu, Lingjia
Chen, Qiang
Wang, Fangfang
Li, Qiuyang
Zeng, Qiang
Huang, Jingcao
Luo, Maowen
Li, Wenxian
Zheng, Yuhuan
Liu, Ting
author_sort Zhao, Dewan
collection PubMed
description BACKGROUND: The physiological approach suggests that an environment associating mesenchymal stromal cells with low O(2) concentration would be most favorable for the maintenance of hematopoietic stem/progenitor cells (HSPCs). To test this hypothesis, we performed a coculture of cord blood CD34(+) cells with Wharton’s jelly mesenchymal stem cells (WJ-MSCs) under different O(2) concentration to simulate the growth of HSPCs in vivo, and assessed the impacts on stemness maintenance and proliferation of cord blood HSPCs in vitro. METHODS: CD34(+) cells derived from cord blood were isolated and cocultured under 1%, 3%, or 20% O(2) concentrations with irradiated WJ-MSCs without adding exogenous cytokines for 7 days. The cultured cells were harvested and analyzed for phenotype and functionality, including total nuclear cells (TNC), CD34(+)Lin(−) cells, colony forming unit (CFU) for committed progenitors, and long-term culture initiating cells (LTC-ICs) for HSPCs. The cytokine levels in the medium were detected with Luminex liquid chips, and the mRNA expression of hypoxia inducible factor (HIF) genes and stem cell signal pathway (Notch, Hedgehog, and Wnt/β-catenin) downstream genes in cord blood HSPCs were confirmed by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: Our results showed that the number of TNC cells, CD34(+)Lin(−) cells, and CFU were higher or similar with 20% O(2) (normoxia) in coculture and compared with 1% O(2) (hypoxia). Interestingly, a 1% O(2) concentration ensured better percentages of CD34(+)Lin(−) cells and LTC-IC cells. The hypoxia tension (1% O(2)) significantly increased vascular endothelial growth factor (VEGF) secretion and decreased interleukin (IL)-6, IL-7, stem cell factor (SCF), and thrombopoietin (TPO) secretion of WJ-MSCs, and selectively activated the Notch, Wnt/β-catenin, and Hedgehog signaling pathway of cord blood HSPCs by HIF-related factors, which may play an important role in stemness preservation and for sustaining HSPC quiescence. CONCLUSIONS: Our data demonstrate that cord blood HSPCs maintain stemness better under hypoxia than normoxia with WJ-MSC coculture, partially due to the increased secretion of VEGF, decreased secretion of IL-6 by WJ-MSCs, and selective activation of stem cell signal pathways in HSPCs. This suggests that the oxygenation may not only be a physiological regulatory factor but also a cell engineering tool in HSPC research, and this may have important translational and clinical implications.
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spelling pubmed-59985412018-06-25 Hypoxia with Wharton’s jelly mesenchymal stem cell coculture maintains stemness of umbilical cord blood-derived CD34(+) cells Zhao, Dewan Liu, Lingjia Chen, Qiang Wang, Fangfang Li, Qiuyang Zeng, Qiang Huang, Jingcao Luo, Maowen Li, Wenxian Zheng, Yuhuan Liu, Ting Stem Cell Res Ther Research BACKGROUND: The physiological approach suggests that an environment associating mesenchymal stromal cells with low O(2) concentration would be most favorable for the maintenance of hematopoietic stem/progenitor cells (HSPCs). To test this hypothesis, we performed a coculture of cord blood CD34(+) cells with Wharton’s jelly mesenchymal stem cells (WJ-MSCs) under different O(2) concentration to simulate the growth of HSPCs in vivo, and assessed the impacts on stemness maintenance and proliferation of cord blood HSPCs in vitro. METHODS: CD34(+) cells derived from cord blood were isolated and cocultured under 1%, 3%, or 20% O(2) concentrations with irradiated WJ-MSCs without adding exogenous cytokines for 7 days. The cultured cells were harvested and analyzed for phenotype and functionality, including total nuclear cells (TNC), CD34(+)Lin(−) cells, colony forming unit (CFU) for committed progenitors, and long-term culture initiating cells (LTC-ICs) for HSPCs. The cytokine levels in the medium were detected with Luminex liquid chips, and the mRNA expression of hypoxia inducible factor (HIF) genes and stem cell signal pathway (Notch, Hedgehog, and Wnt/β-catenin) downstream genes in cord blood HSPCs were confirmed by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: Our results showed that the number of TNC cells, CD34(+)Lin(−) cells, and CFU were higher or similar with 20% O(2) (normoxia) in coculture and compared with 1% O(2) (hypoxia). Interestingly, a 1% O(2) concentration ensured better percentages of CD34(+)Lin(−) cells and LTC-IC cells. The hypoxia tension (1% O(2)) significantly increased vascular endothelial growth factor (VEGF) secretion and decreased interleukin (IL)-6, IL-7, stem cell factor (SCF), and thrombopoietin (TPO) secretion of WJ-MSCs, and selectively activated the Notch, Wnt/β-catenin, and Hedgehog signaling pathway of cord blood HSPCs by HIF-related factors, which may play an important role in stemness preservation and for sustaining HSPC quiescence. CONCLUSIONS: Our data demonstrate that cord blood HSPCs maintain stemness better under hypoxia than normoxia with WJ-MSC coculture, partially due to the increased secretion of VEGF, decreased secretion of IL-6 by WJ-MSCs, and selective activation of stem cell signal pathways in HSPCs. This suggests that the oxygenation may not only be a physiological regulatory factor but also a cell engineering tool in HSPC research, and this may have important translational and clinical implications. BioMed Central 2018-06-13 /pmc/articles/PMC5998541/ /pubmed/29895317 http://dx.doi.org/10.1186/s13287-018-0902-5 Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Zhao, Dewan
Liu, Lingjia
Chen, Qiang
Wang, Fangfang
Li, Qiuyang
Zeng, Qiang
Huang, Jingcao
Luo, Maowen
Li, Wenxian
Zheng, Yuhuan
Liu, Ting
Hypoxia with Wharton’s jelly mesenchymal stem cell coculture maintains stemness of umbilical cord blood-derived CD34(+) cells
title Hypoxia with Wharton’s jelly mesenchymal stem cell coculture maintains stemness of umbilical cord blood-derived CD34(+) cells
title_full Hypoxia with Wharton’s jelly mesenchymal stem cell coculture maintains stemness of umbilical cord blood-derived CD34(+) cells
title_fullStr Hypoxia with Wharton’s jelly mesenchymal stem cell coculture maintains stemness of umbilical cord blood-derived CD34(+) cells
title_full_unstemmed Hypoxia with Wharton’s jelly mesenchymal stem cell coculture maintains stemness of umbilical cord blood-derived CD34(+) cells
title_short Hypoxia with Wharton’s jelly mesenchymal stem cell coculture maintains stemness of umbilical cord blood-derived CD34(+) cells
title_sort hypoxia with wharton’s jelly mesenchymal stem cell coculture maintains stemness of umbilical cord blood-derived cd34(+) cells
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5998541/
https://www.ncbi.nlm.nih.gov/pubmed/29895317
http://dx.doi.org/10.1186/s13287-018-0902-5
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