Lv Cheng-Xiao, Duan Hua, Wang Sha, Gan Lu, Xu Qian
Department of Minimally Invasive Gynecologic Center, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, People's Republic of China.
Reprod Sci. 2020 Jun;27(6):1372-1381. doi: 10.1007/s43032-020-00165-y. Epub 2020 Jan 31.
Umbilical cord mesenchymal stem cells (UCMSCs) have been proposed as an ideal source for cell-based therapy to promote endometrial repair and regeneration. Furthermore, increasing evidence has indicated that UCMSC-derived exosomes (UCMSC-exos) act as important paracrine mediators to recapitulate the features of MSCs and may play a vital role in this process. UCMSCs and human endometrial stromal cells (ESCs) were isolated and characterized. ESCs were cocultured with UCMSCs and further assessed by flow cytometry and EdU incorporation assays. UCMSC-exos were extracted by differential ultracentrifugation and identified by western blots, transmission electron microscopy, and nanoparticle tracking analysis. The internalization of UCMSC-exos by ESCs was observed under a confocal microscope. ESCs were treated with UCMSC-exos at different concentrations and for different durations, with cell viability evaluated by CCK-8 assays. The cell cycle analysis showed that the percentage of ESCs in S phase significantly increased after coculture with UCMSCs, whereas it significantly decreased after inhibition of UCMSC-exo secretions. EdU incorporation assays also showed a similar trend. The isolated UCMSC-exos had a typical cup-shaped morphology with a monolayer membrane, expressed the specific exosomal markers Alix, CD63, and TSG101 and were approximately 60 to 200 nm in diameter. The PKH26-labeled UCMSC-exos were incorporated into ESCs. Moreover, UCMSC-exos enhanced the cell growth and viability of ESCs in a dose-dependent manner, and the effects occurred in a short period of time. UCMSC-exos promote the proliferation of ESCs in a dose-dependent manner; thus, they could be used as a potential treatment to promote endometrial repair.
脐带间充质干细胞（UCMSCs）已被认为是基于细胞的疗法促进子宫内膜修复和再生的理想来源。此外，越来越多的证据表明，UCMSC衍生的外泌体（UCMSC-exos）作为重要的旁分泌介质，重现了间充质干细胞的特征，并且可能在此过程中发挥至关重要的作用。分离并鉴定了UCMSCs和人子宫内膜基质细胞（ESCs）。将ESCs与UCMSCs共培养，并通过流式细胞术和EdU掺入试验进一步评估。通过差速超速离心法提取UCMSC-exos，并通过蛋白质免疫印迹、透射电子显微镜和纳米颗粒跟踪分析进行鉴定。在共聚焦显微镜下观察ESCs对UCMSC-exos的内化情况。用不同浓度和不同持续时间的UCMSC-exos处理ESCs，通过CCK-8试验评估细胞活力。细胞周期分析表明，与UCMSCs共培养后，ESCs处于S期的百分比显著增加，而在抑制UCMSC-exo分泌后显著降低。EdU掺入试验也显示出类似的趋势。分离出的UCMSC-exos具有典型的杯状形态，有单层膜，表达特异性外泌体标志物Alix、CD63和TSG101，直径约为60至200nm。PKH26标记的UCMSC-exos被ESCs摄取。此外，UCMSC-exos以剂量依赖的方式增强了ESCs的细胞生长和活力，且这些作用在短时间内即可出现。UCMSC-exos以剂量依赖的方式促进ESCs的增殖；因此，它们可作为促进子宫内膜修复的潜在治疗方法。
