组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2023, Vol. 19 ›› Issue (2): 111-.

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两种消化液传代培养的人脂肪干细胞生物学特性比较研究

  

  • 出版日期:2023-04-01 发布日期:2023-05-09

Comparative study on biological characteristics of human adipose stem cells cultured in two kinds of digestion reagents

  • Online:2023-04-01 Published:2023-05-09

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摘要：

目的　比较两种消化试剂传代对人脂肪干细胞生物学特性的影响，探寻可替代胰蛋白酶的细胞消化试剂。
方法　采用类胰蛋白酶（TrypLE Express）与胰蛋白酶（Trypsin）两种细胞消化试剂，对原代提取的人脂肪干细胞生
物学特性及成脂分化能力进行比较。结果　第三代脂肪干细胞在 TrypLETM Express 与 Trypsin 试剂消化条件下活细胞
百分比无显著性差异（P>0.05）。流式细胞仪检测结果表明，CD90、CD73、CD105 等细胞表面抗原标志物呈现阳性
表达，CD11b、CD34、CD19、CD45、HLA-DR 等呈现阴性表达，两组消化试剂作用下阳性细胞的百分比无显著性
差异（P>0.05）。成脂诱导细胞脂滴染色结果用 Imagepro plus 进行半定量分析，两组消化试剂作用下成脂分化率无显
著性差异（P>0.05），接种第 1~7 天细胞增殖能力无明显差异（P>0.05）。结论　两种消化试剂作用对人脂肪干细胞
生物学特性的影响无显著性差异，TrypLE Express 可完全代替 Trypsin 用于脂肪干细胞体外传代培养。

关键词:

Abstract:

Objective To compare the effects of two kinds of digestion reagents on the biological characteristics of human
adipose derived stem cells, and to explore cell digestion reagents that can replace trypsin. Methods Two cell digestion re
agents (TrypLE Express and Trypsin) were used to compare the biological characteristics and adipogenic differentiation ability
of primary human adipose stem cells. Results　There was no significant difference in the percentage of viable cells in the third
generation of adipose stem cells digested with TrypLETM Express and Trypsin (P>0.05). The results of flow cytometry showed that
CD90, CD73, CD105 and other cell surface antigen markers were positively expressed, while CD11b, CD34, CD19, CD45, HLA
DR were negatively expressed. There was no significant difference in the percentage of positive cells between the two groups un
der the treatment of digestive reagents (P>0.05). Imagepro Plus was used for semi-quantitative analysis of lipid droplet staining
results of adipogenic induced cells. There was no significant difference in adipogenic differentiation rate between the two groups
under the action of digestive reagents (P>0.05), and no significant difference in cell proliferation ability from the first day to the
seventh day of inoculation (P>0.05). Conclusion　There was no significant difference between the two digestion reagents on the
biological characteristics of human adipose stem cells. TrypLE Express could completely replace Trypsin for the culture of adi
pose stem cells in vitro

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