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中华损伤与修复杂志(电子版) ›› 2023, Vol. 18 ›› Issue (03) : 229 -234. doi: 10.3877/cma.j.issn.1673-9450.2023.03.009

论著

对人脐带间充质干细胞外泌体影响热损伤人皮肤成纤维细胞迁移的分析
黄瑞娟, 德奇, 巴特(), 周彪   
  1. 014010 包头,内蒙古包钢医院
    010110 呼和浩特,内蒙古医科大学第三临床医学院
  • 收稿日期:2022-07-01 出版日期:2023-06-01
  • 通信作者: 巴特
  • 基金资助:
    重大疾病防治科技行动计划(2018-ZX-01S-001); 内蒙古自治区自然科学基金项目(2020MS08022); 内蒙古自治区自然科学基金项目(2020MS08181); 内蒙古医科大学科技百万工程(联合)项目(YKD2016KJBW(LH)041); 内蒙古自治区科技创新引导项目(XCYD2020BT03)

Influence of human umbilical cord mesenchymal stem cell exosomes on migration of heat-damaged human skin fibroblasts

Ruijuan Huang, Qi De, Te Ba(), Biao Zhou   

  1. Laser Treatment Center, Inner Mongolia Baogang Hospital, Baotou 014010, China
    The Third Clinical Medical College of Inner Mongolia Medical University, Huhhot 010110, China
  • Received:2022-07-01 Published:2023-06-01
  • Corresponding author: Te Ba
引用本文:

黄瑞娟, 德奇, 巴特, 周彪. 对人脐带间充质干细胞外泌体影响热损伤人皮肤成纤维细胞迁移的分析[J]. 中华损伤与修复杂志(电子版), 2023, 18(03): 229-234.

Ruijuan Huang, Qi De, Te Ba, Biao Zhou. Influence of human umbilical cord mesenchymal stem cell exosomes on migration of heat-damaged human skin fibroblasts[J]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2023, 18(03): 229-234.

目的

初步探讨人脐带间充质干细胞外泌体对热损伤人皮肤成纤维细胞迁移的影响。

方法

分离脐带间充质干细胞(hucMSCs)并进行形态学鉴定,流式细胞术检测细胞表面标志物。采用PEG沉淀法联合差速离心法获取人脐带间充质干细胞外泌体(hucMSC-Exos)并鉴定。分离培养出人真皮成纤维细胞,建立成纤维细胞热损伤模型。将热损伤成纤维细胞分为3组,分别加入不同剂量(0 μg、50 μg、100 μg)hucMSC-Exos溶液,依次进行细胞划痕、Transwell实验,观察hucMSC-Exos对热损伤成纤维细胞迁移的影响。

结果

24 h后,50 μg和100 μg hucMSC-Exos组热损伤成纤维细胞的划痕愈合率分别为42.86%±6.54%、55.42%±8.53%,均大于0 μg hucMSC-Exos组(23.21%±8.25%),差异有统计学意义(t=-7.111、-9.268,P<0.05)。24 h后,50 μg、100 μg hucMSC-Exos组热损伤成纤维细胞迁移个数分别为(31.47±4.31)、(39.53±3.11)个,相比于0 μg hucMSC-Exos组[(19.20±2.91)个]显著增多,组间比较差异有统计学意义(t=-10.213、-28.710,P<0.05)。

结论

人脐带间充质干细胞外泌体能够促进热损伤人皮肤成纤维细胞迁移。

Objective

To preliminarily investigate the effect of human umbilical cord mesenchymal stem cell exosomes on the migration of heat-damaged human skin fibroblasts.

Methods

Umbilical cord mesenchymal stem cells (hucMSCs) were isolated and morphologically identified surface markers by flow cytometry. Human umbilical cord mesenchymal stem cell exosomes (hucMSC-Exos) were obtained and identified by PEG precipitation combined with differential centrifugation. Human dermal fibroblasts were isolated and cultured, and a model of fibroblast thermal injury was established. Heat-damaged fibroblasts were divided into three groups, different doses of hucMSC-Exos solution (0 μg, 50 μg, 100 μg) were added, and cell scratches and Transwell experiments were performed sequentially to observe the effect of hucMSC-Exos on the migration of heat-damaged fibroblasts.

Results

After 24 h, the scratch healing rates of heat-damaged fibroblasts in 50 μg and 100 μg hucMSC-Exos groups were 42.86%±6.54% and 55.42%±8.53%, respectively, which were higher than that in 0μg hucMSC-Exos group (23.21%±8.25%) with statistically differences (t=-7.111、-9.268, P<0.05). After 24 h, the migration numbers of heat-damaged fibroblasts in 50 μg and 100 μg hucMSC-Exos groups were 31.47±4.31 and 39.53±3.11, respectively, which were more than that in 0 μg hucMSC-Exos group (19.20±2.91), the differences were statistically significant (t=-10.213、-28.710, P<0.05).

Conclusion

Human umbilical cord mesenchymal stem cell exosomes can promote the migration of heat-damaged human skin fibroblasts.

图1 脐带间充质干细胞、外泌体、成纤维细胞培养及鉴定。A示脐带间充质干细胞流式细胞仪鉴定;B示脐带间充质干细胞传代培养(100×);C示外泌体透射透射电子显微镜观察;D示外泌体粒径分析;E示成纤维细胞传代培养(50×);F示成纤维细胞波形蛋白染色(100×)
图2 细胞热损伤模型建立及评估。A示细胞划痕实验(50×);B示Transwell实验(100×)
图3 hucMSC-Exos对热损伤成纤维细胞迁移的影响。A示细胞划痕实验(50×); B示Transwell实验(100×)
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