江苏大学学报（医学版）2026，Vol.36Issue(2)：129-135,7.DOI:10.13312/j.issn.1671-7783.y250096
反复冻融与超声波处理制备的干细胞裂解液修复皮肤损伤的效果比较
Comparison of the effects of stem cell lysate prepared by repeated freeze-thaw and ultrasonic treatment on skin injury repair
摘要
Abstract
Objective:To compare the effects of stem cell lysate prepared by repeated freeze-thaw method and ultrasonic disruption method on skin injury repair,and screen the optimal preparation method of stem cell lysate.Methods:Human skin fibroblasts were resuspended in normal saline at four densities(1×105/mL,1×106/mL,5×106/mL,1×107/mL),and treated by repeated freeze-thaw and ultrasonic disruption,respectively.The cell disruption rate and protein content were detected by BCA assay to screen the optimal preparation conditions of the two methods.The scratch healing ability,viability and migration number of human skin fibroblasts were detected by cell scratch healing assay,CCK-8 method and Transwell assay,respectively.The protein concentration of epidermal growth factor(EGF),vascular endothelial growth factor(VEGF)and stem cell factor(SCF)in the lysate were detected by ELISA.Results:The optimal condition of ultrasonic disruption method was cell concentration of 5×106/mL with ultrasonic treatment for 5 min;the optimal condition of repeated freeze-thaw method was cell concentration of 5×106/mL with three freeze-thaw cycles(45 min).There was no significant difference in cell disruption rate between the two methods(P＞0.05),and the total protein content of lysate prepared by ultrasonic disruption method was higher.There was no significant difference in the effect of lysates prepared by the two methods on the viability of human skin fibroblasts(P＞0.05).Compared with ultrasonic disruption method,the lysate prepared by repeated freeze-thaw method could significantly promote the migration of human skin fibroblasts and improve the efficiency of cell scratch repair(P＜0.05).ELISA results showed that the concentrations of VEGF and SCF proteins in the lysate prepared by repeated freeze-thaw method were significantly higher than those by ultrasonic disruption method(P＜0.05 or P＜0.01),and there was no significant difference in EGF content(P＞0.05).Conclusion:The stem cell lysate prepared by repeated freeze-thaw method has a better effect on skin injury repair than that by ultrasonic disruption method.关键词
皮肤损伤修复/干细胞裂解液/反复冻融法/超声波破碎法/成纤维细胞Key words
skin injury repair/stem cell lysate/repeated freeze-thaw method/ultrasonic disruption method/fibroblast分类
医药卫生引用本文复制引用
李凯,徐聪,秦金伟..反复冻融与超声波处理制备的干细胞裂解液修复皮肤损伤的效果比较[J].江苏大学学报（医学版）,2026,36(2):129-135,7.基金项目
中国博士后科学基金资助项目(2023MZ32157) （2023MZ32157）
