Paracrine abilities of adipose-derived mesenchymal stem cells and umbilical cord mesenchymal stem cells under different oxygen concentrations

doi:10.3969/j.issn.2095-4344.0880

Abstract

Abstract:

BACKGROUND: Therapeutic mechanisms of mesenchymal stem cells for acute myocardial infarction are related to the migration, colonization, and differentiation of mesenchymal stem cells, and more importantly, it is related to the paracrine effects of mesenchymal stem cells.
OBJECTIVE: To evaluate the paracrine and vascular regeneration abilities of human adipose-derived mesenchymal stem cells (hAD-MSCs) and human umbilical cord mesenchymal stem cells (hUC-MSCs) under low oxygen concentrations, thereby providing the basis for the selection of appropriate mesenchymal stem cells used in the treatment of acute myocardial infarction.
METHODS: hUC-MSCs and hAD-MSCs were isolated and cultured, and cell immunophenotype was identified using flow cytometry. Two kinds of cells were pretreated with 3%, 5% and 21% (control) O₂ for 24 hours. Enzyme linked immunosorbent assay was used to detect the contents of hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), nerve growth factor (NGF) and keratinocyte growth factor (KGF) in the cell supernatant in each group.
RESULTS AND CONCLUSION: hUC-MSCs and hAD-MSCs were positive for CD73, CD90 and CD105, and negative for CD34, CD45, CD54 and HLA-DR. Under different oxygen concentrations, the VEGF level in the hAD-MSCs was significantly higher than that in the hUC-MSCs (P < 0.01). Under 3% O₂ conditions, the HGF level in the hAD-MSCs was significantly higher than that in the hUC-MSCs (P < 0.05). Under 3% and 5% O₂, the NGF level in the hUC-MSCs was significantly higher than that in the hAD-MSCs (P < 0.01). Under 3% and 5% O₂, the KGF level in the hUC-MSCs was significantly higher than that in the hAD-MSCs (P < 0.01 or P < 0.05). Under 21% O₂, the KGF level in the hAD-MSCs was significantly higher than that in the hUC-MSCs (P < 0.01). To conclude, hypoxia can promote the paracrine effects of and hAD-MSCs. Moreover, hUC-MSCs show better paracrine ability than hAD-MSCs. Therefore, hAD-MSCs are more likely to be a stem cell therapy suitable for myocardial infarction.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Myocardial Infarction, Adipose Tissue, Umbilical Cord, Mesenchymal Stem Cells, Cell Hypoxia, Vascular Endothelial Growth Factors, Hepatocyte Growth Factor, Nerve Growth Factor, Tissue Engineering

CLC Number:

R394.2

Yang Ji-nong, Jiang Yi-yao, Yuan Chao, Liu Zhi-gang, Liu Xiao-cheng. Paracrine abilities of adipose-derived mesenchymal stem cells and umbilical cord mesenchymal stem cells under different oxygen concentrations[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(21): 3322-3327.

Figures/Tables 1

2.1 脂肪间充质干细胞与脐带间充质干细胞的形态原代细胞在24-48 h内，少量细胞开始贴壁，呈梭形，多角形分布；5-7 d可见贴壁细胞数量增多；7-10 d融合率达80%以上，12 d左右细胞密度增高。按1∶2-1∶3的比例传代，传代细胞于接种后4-6 h迅速贴壁，最初为短粗的梭形或卵圆形。随后，细胞展开呈梭形、多角形，细胞均匀分布，可呈片状融合，低倍镜下细胞排列紧密，具有方向性，杂质细胞明显减少。经传代扩增，细胞形态不变(图1)。 2.2 脂肪间充质干细胞与脐带间充质干细胞的免疫表型鉴定结果脂肪间充质干细胞与脐带间充质干细胞的免疫表型表达具有共性，即CD73、CD90、CD105呈阳性表达，而CD34、CD45、CD54、HLA-DR为阴性表达。 2.3 脂肪间充质干细胞与脐带间充质干细胞的肝细胞生长因子、血管内皮生长因子、神经生长因子、角质细胞生长因子蛋白分泌量在低氧环境下，2种干细胞的血管内皮生长因子分泌均呈现增高趋势。在脂肪间充质干细胞中，与对照组相比，体积分数为3% O2预处理组血管内皮生长因子蛋白分泌量差异有非常显著性意义(P < 0.01)。在脐带间充质干细胞中，与对照组相比，体积分数为3% O2预处理组血管内皮生长因子蛋白分泌量同样差异有非常显著性意义(P < 0.01)。在体积分数为3% O2条件下，脂肪间充质干细胞与脐带间充质干细胞的血管内皮生长因子蛋白分泌量差异有非常显著性意义(P < 0.01)，在体积分数为5% O2条件下，脂肪间充质干细胞与脐带间充质干细胞的血管内皮生长因子蛋白分泌量差异有非常显著性意义(P < 0.01)，见表1，图2A。在低氧环境下，2种干细胞的肝细胞生长因子分泌均呈现增高趋势。在脂肪间充质干细胞中，与对照组相比，体积分数为3% O2预处理组肝细胞生长因子蛋白分泌量差异有非常显著性意义(P < 0.01)。在脐带间充质干细胞中，与对照组相比，体积分数为3% O2预处理组肝细胞生长因子蛋白分泌量同样差异有非常显著性意义(P < 0.01)。在体积分数为5% O2条件下，脂肪间充质干细胞与脐带间充质干细胞的肝细胞生长因子蛋白分泌量差异有显著性意义(P < 0.05)，见表2，图2B。在低氧环境下，脐带间充质干细胞的神经生长因子分泌呈现增高趋势，与对照组相比，体积分数为3% O2预处理组神经生长因子蛋白分泌量差异有非常显著性意义(P < 0.01)。在体积分数为3% O2条件下，脂肪间充质干细胞与脐带间充质干细胞的神经生长因子蛋白分泌量差异有非常显著性意义(P < 0.01)，在体积分数为5% O2条件下，脂肪间充质干细胞与脐带间充质干细胞的神经生长因子蛋白分泌量差异有非常显著性意义(P < 0.01)，见表3，图2C。在低氧环境下，脐带间充质干细胞的角质细胞生长因子分泌呈现增高趋势，与对照组相比，体积分数为3% O2预处理组角质细胞生长因子蛋白分泌量差异有非常显著性意义(P < 0.01)。在体积分数为3% O2条件下，脂肪间充质干细胞与脐带间充质干细胞的角质细胞生长因子蛋白分泌量差异有非常显著性意义(P < 0.01)，在体积分数为5% O2条件下，脂肪间充质干细胞与脐带间充质干细胞的角质细胞生长因子蛋白分泌量差异有显著性意义(P < 0.05)，见表4，图2D。"

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