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. 2023 Feb;31(1):227-232.
doi: 10.19746/j.cnki.issn.1009-2137.2023.01.036.

[Effect of Hypoxia-Supported Umbilical Cord Mesenchymal Stem Cells on the Expansion of Cord Blood Mononuclear Cells in vitro]

[Article in Chinese]
Jing Yu  1 , Li Xu  2
Affiliations

[Effect of Hypoxia-Supported Umbilical Cord Mesenchymal Stem Cells on the Expansion of Cord Blood Mononuclear Cells in vitro]

[Article in Chinese]
Jing Yu et al. Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023 Feb.

Abstract
in English, Chinese

Objective: To explore the effect of hypoxia-supported umbilical cord mesenchymal stem cell (UC-MSC) on the expansion of cord blood mononuclear cell （MNC） in vitro.

Methods: The isolated cord blood mononuclear cells were inoculated on the preestablished umbilical cord mesenchymal stem cell layer and cultured under hypoxic conditions (3％ O2) and the experimental groups were normoxia (MNCs were cultured under normoxic conditions), hypoxia (MNCs were cultured under hypoxic conditions), UC-MSC (MNCs were cultured with UC-MSC under normoxic conditions), and UC-MSC+hypoxia (MNCs were cultured with UC-MSC under hypoxic conditions). To further investigate the combinational effect of 3 factors of SCF+FL+TPO (SFT) on expansion of cord blood MNCs in vitro in hypoxia-supported UC-MSC culture system, the experiments were further divided into group A (MNCs were cultured with UC-MSC and SFT under normoxic conditions), group B (MNCs were cultured with UC-MSC under hypoxic conditions), group C (MNCs were cultured with UC-MSC and SFT under hypoxic conditions). The number of nucleated cells (TNC), CD34+ cell, CFU and CD34+CXCR4+, CD34+CD49d+, CD34+CD62L+ cells of each groups were detected at 0, 7, 10 and 14 days, respectively.

Results: Compared with group hypoxia and UC-MSC, group UC-MSC+hypoxia effectively promoted the expansion of TNC, CD34+ cell and CFU, and upregulated the expression level of adhesion molecule and CxCR4 of the cord blood CD34+ cell（P<0.05）. After culturing for 14 days, compared with group A and group B, group C effectively promoted the expansion of cord blood MNC at different time points（P<0.05）, and the effect of group A was better than that of group B at 7 and 10 days（P<0.05）.

Conclusion: Hypoxia-supported UC-MSC efficiently promoted the expansion and expression of adhesion molecule and CXCR4 of cord blood CD34+ cell, and the effect of expansion could be enhanced when SFT 3 factors were added.

题目: 低氧支持的脐带间充质干细胞对脐血单个核细胞体外扩增的影响.

目的: 探讨低氧支持的脐带间充质干细胞（UC-MSC）对人脐血单个核细胞的体外扩增的影响.

方法: 将分离的脐血单个核细胞（MNC）接种在预先建立的UC-MSC层上，在低氧（3％ O2) 条件下分组培养，实验分组为常氧（常氧+脐血MNC）、低氧（低氧+脐血MNC）、UC-MSC（常氧+UC-MSC+脐血MNC）、UC-MSC+低氧（低氧+UC-MSC+脐血MNC）共4组。为进一步探讨SCF+FL+TPO（SFT） 3种因子组合在低氧支持的UC-MSC培养体系中脐血MNC体外扩增的作用，实验进一步分组为A（常氧+UC-MSC+SFT+脐血MNC）、B组（低氧+UC-MSC+脐血MNC）、C（低氧+UC-MSC+SFT+脐血MNC）共3组，培养0、7、10、14 d检测各组总有核细胞数（TNC）、 CD34+细胞数、集落形成单位(CFU)数及CD34+CXCR4+、CD34+CD49d+、CD34+CD62L+细胞数并进行比较分析.

结果: UC-MSC+低氧组较低氧组、UC-MSC组能有效地扩增脐血TNC、CD34+细胞、CFU数（P<0.05）, 并可明显提高脐血CD34+细胞上黏附分子和CXCR4的表达（P<0.05）；经过14 d的培养，C组各时间点检测指标水平均明显高于A、B组（P<0.05），A组7、10 d扩增效果明显优于B组（P<0.05）.

结论: 低氧支持的UC-MSC可有效扩增脐血造血细胞，加入SFT因子组合可明显扩增脐血MNC并能提高脐血CD34+细胞上黏附分子和CXCR4的表达.

Keywords: cord blood; exvivo expansion; hypoxia; mononuclear cell; umbilical cord mesenchymal stem cell.

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