不同胎牛血清浓度培养的人脐带间充质干细胞的条件培养基对角膜上皮细胞损伤修复的影响  

Effects of human umbilical cord mesenchymal stem cell conditioned medium with different fetal bovine serum concentrations on damage and repair of human corneal epithelial cells

作　　者：潘华婴 赵成海[1] 魏文斌[2] Pan Huaying;Zhao Chenghai;Wei Wenbin(Department of Pathophysiology,China Medical University,Shenyang110001,China;不详)

机构地区：[1]中国医科大学病理生理学教研室,沈阳110001 [2]北京同仁医院眼科中心

出　　处：《中国药物与临床》2021年第6期917-920,共4页Chinese Remedies & Clinics

摘　　要：目的探讨不同胎牛血清(FBS)浓度培养下人脐带间充质干细胞(hUCMSCs)条件培养液(hUCMSCCM)对角膜上皮细胞损伤修复的促进作用。方法采用贴壁法培养,以含有1%,3%,5%,10%的FBS培养基培养P6代hUCMSCs并收集4种培养条件下的培养基,利用CCK-8方法检测细胞hUCMSCCM对角膜上皮细胞增殖的影响;收集4种hUCMSCCM培养损伤的角膜上皮细胞,检测其对角膜上皮细胞损伤修复的影响;采用酶联免疫吸附试验(ELISA)检测4种hUCMSCCM中血小板源性生长因子(PDGF)和白细胞介素-8(IL-8)的含量。结果①收集的4种hUCMSCCM均能促进角膜上皮细胞增殖,随着培养时间的延长,其中1%hUCM-SCCM和3%hUCMSCCM2组条件下,角膜上皮细胞的增殖能力显著高于其他实验组和对照组;②4种hUCM-SCCM均促进角膜上皮细胞的迁移,1%hUCMSCCM在24 h促进角膜上皮细胞损伤修复最为显著,随着时间延长,除10%hUCMSCCM组外其余组均表现出显著的促进角膜上皮细胞损伤修复的能力;③4种UCMSCCM中PDGF的分泌具有差异性,随着血清浓度的增大,PDGF的含量逐渐减少;而IL-8的分泌与血清浓度的增加呈现正相关性。结论不同浓度FBS培养的hUCMSCCM对角膜上皮细胞增殖和损伤修复具有促进作用,其中1%FBS hUCMSCCM在24 h修复和促增殖最为显著;其作用是通过分泌修复因子促进角膜上皮细胞的增殖和迁移,从而保护角膜上皮细胞。干细胞条件培养基将成为角膜上皮损伤的新治疗方法。Objective To investigate the promotion of human umbilical cord mesenchymal stem cell conditioned medium(hUCMSCCM)with different fetal bovine serum(FBS)levels on the damage and repair of human corneal epithelial cells(HCECs).Methods Adherence method was used to culture passage 6(P6)hUCMSCs with the medium containing different FBS levels(1%,3%,5%and 10%).The hUCMSCCMs under 4 culture conditions were collected.The CCK-8 assay was used to determine the effect of hUCMSCCM on the proliferation of HCECs.The damaged HCECs were cultured by 4 hUCMSCCMs,and the effects on the repair of HCECs damage were examined.Enzyme-linked immunoassay(ELISA)was used to determine the levels of platelet-derived growth factor(PDGF)and interleukin-8(IL-8)in the 4 hUCMSCCMs.Results①The 4 hUCMSCCMs could promote the proliferation of HCECs.With the extension of the culture time,the proliferation ability of HCECs in the 1%FBS hUCMSCCM group and 3%FBS hUCMSCCM group was significantly higher than that in the other study groups and control group.②All the 4 hUCMSCCMs could promote the migration of HCECs.1%FBS hUCMSCCM showed the most significantly promotion on the repair of HCECs damage at 24 h.With the extension of the culture time,all groups except the 10%FBS hUCMSCCM group showed significant ability to promote the repair of HCECs damage.③There was difference in the secretion of PDGF among the four hUCMSCCM groups.With the increase of serum concentration,the content of the PDGF level gradually decreased along with the increasing serum level.The secretion of IL-8 was positively correlated with the increase of serum level.Conclusion The hUCMSCCM cultured with different FBS levels may promote the proliferation and damage repair of HCECs.Among them,1%FBS hUCMSCCM shows the most significantly repair and promotion of proliferation at 24 hours.Its role is to promote the proliferation and migration of HCECs by secreting repair factors,and thereby to protect HCECs.Stem cell conditioned medium will become a new treatment method for damage of HCECs.

关 键 词：胎牛血清 人脐带间充质干细胞 培养基 条件性 角膜上皮细胞 损伤修复 

分 类 号：R772.2[医药卫生—眼科]