Yu Jing, Xu Li
Medical College, Anhui University of Science and Technology, Huainan 232001, Anhui Province, China.
Anhui No.2 Provincial People's Hospital, Hefei 230041, Anhui Province, China,E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023 Feb;31(1):227-232. doi: 10.19746/j.cnki.issn.1009-2137.2023.01.036.
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.
The isolated cord blood mononuclear cells were inoculated on the preestablished umbilical cord mesenchymal stem cell layer and cultured under hypoxic conditions (3% O) 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 CD34CXCR4, CD34CD49d, CD34CD62L cells of each groups were detected at 0, 7, 10 and 14 days, respectively.
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(<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(<0.05), and the effect of group A was better than that of group B at 7 and 10 days(<0.05).
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.
探讨低氧支持下的脐带间充质干细胞(UC-MSC)对脐血单个核细胞(MNC)体外扩增的影响。
将分离的脐血单个核细胞接种于预先建立的脐带间充质干细胞层上,在低氧条件(3%O₂)下培养,实验组分别为常氧组(MNC在常氧条件下培养)、低氧组(MNC在低氧条件下培养)、UC-MSC组(MNC与UC-MSC在常氧条件下共培养)和UC-MSC+低氧组(MNC与UC-MSC在低氧条件下共培养)。为进一步研究干细胞因子(SCF)+促血小板生成素(TPO)+白细胞介素-3(FL)(SFT)这3种因子在低氧支持的UC-MSC培养体系中对脐血MNC体外扩增的联合作用,实验进一步分为A组(MNC与UC-MSC和SFT在常氧条件下共培养)、B组(MNC与UC-MSC在低氧条件下共培养)、C组(MNC与UC-MSC和SFT在低氧条件下共培养)。分别在0、7、10和14天检测各组有核细胞(TNC)、CD34⁺细胞、集落形成单位(CFU)以及CD34⁺CXCR4⁺、CD34⁺CD49d⁺、CD34⁺CD62L⁺细胞的数量。
与低氧组和UC-MSC组相比,UC-MSC+低氧组能有效促进TNC、CD34⁺细胞和CFU的扩增,并上调脐血CD34⁺细胞黏附分子和CxCR4的表达水平(P<0.05)。培养14天后,与A组和B组相比,C组在不同时间点均能有效促进脐血MNC的扩增(P<0.05),且A组在7天和10天时的作用优于B组(P<0.05)。
低氧支持下的UC-MSC能有效促进脐血CD34⁺细胞的扩增及其黏附分子和CXCR4的表达,添加SFT 3种因子可增强扩增效果。