Yi Dan, Zhang Yong-Yi, Jiang Wen-Li, Li Mo-Lin, Chen Xiang-Hui, Yu Jiang, Yi Hong-Yu, Zhu Ya-Qiong, Wang Yue-Xiang
Medical School of Chinese PLA,Beijing 100853,China.
Department of Ultrasound,the First Medical Centre,Chinese PLA General Hospital,Beijing 100853,China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2023 Jun;45(3):374-381. doi: 10.3881/j.issn.1000-503X.15332.
Objective To investigate the effect of human platelet-rich plasma-derived exosomes(PRP-exos)on the proliferation of Schwann cell(SC)cultured . Methods PRP-exos were extracted by polymerization-precipitation combined with ultracentrifugation.The morphology of PRP-exos was observed by transmission electron microscopy,and the concentration and particle size distribution of PRP-exos were determined by nanoparticle tracking analysis.Western blotting was employed to determine the expression of the marker proteins CD63,CD81,and CD9 on exosome surface and the platelet membrane glycoprotein CD41.The SCs of rats were isolated and cultured,and the expression of the SC marker S100β was detected by immunofluorescence staining.The fluorescently labeled PRP-exos were co-cultured with SCs for observation of their interaction.EdU assay was employed to detect the effect of PRP-exos on SC proliferation,and CCK-8 assay to detect the effects of PRP-exos at different concentrations(0,10,20,40,80,and 160 μg/ml)on SC proliferation. Results The extracted PRP-exos appeared as uniform saucer-shaped vesicles with the average particle size of(122.8±38.7)nm and the concentration of 3.5×10 particles/ml.CD63,CD81,CD9,and CD41 were highly expressed on PRP-exos surface(<0.001,=0.025,=0.004,and =0.032).The isolated SCs expressed S100β,and PRP-exos could be taken up by SCs.PRP-exos of 40,80,and 160 μg/ml promoted the proliferation of SCs,and that of 40 μg/ml showed the best performance(all <0.01). Conclusions High concentrations of PRP-exos can be extracted from PRP.PRP-exos can be taken up by SCs and promote the proliferation of SCs cultured .
目的 探讨人富血小板血浆来源外泌体(PRP-exos)对培养的雪旺细胞(SC)增殖的影响。方法 采用聚合法沉淀结合超速离心法提取PRP-exos。通过透射电子显微镜观察PRP-exos的形态,采用纳米颗粒跟踪分析测定PRP-exos的浓度和粒径分布。采用蛋白质免疫印迹法检测外泌体表面标志物蛋白CD63、CD81和CD9以及血小板膜糖蛋白CD41的表达。分离培养大鼠SCs,通过免疫荧光染色检测SC标志物S100β的表达。将荧光标记的PRP-exos与SCs共培养以观察它们之间的相互作用。采用EdU法检测PRP-exos对SC增殖的影响,采用CCK-8法检测不同浓度(0、10、20、40、80和160 μg/ml)的PRP-exos对SC增殖的影响。结果 提取的PRP-exos呈均匀的碟形囊泡,平均粒径为(122.8±38.7)nm,浓度为3.5×10颗粒/ml。PRP-exos表面CD63、CD81、CD9和CD41高表达(<0.001,=0.025,=0.004,=0.032)。分离的SCs表达S100β,PRP-exos可被SCs摄取。40、80和160 μg/ml的PRP-exos促进SCs增殖,40 μg/ml的效果最佳(均<0.01)。结论 可从PRP中提取高浓度的PRP-exos。PRP-exos可被SCs摄取并促进培养的SCs增殖。