Application of Serum Starvation Strategy in the Treatment of Canine(Canis lupus familiaris)Diabetes with Mesenchymal Stem Cells  

血清饥饿法在间充质干细胞治疗犬糖尿病中的应用

作　　者：DU Jia-Xin LI Ba-Lun KOU Zheng WU Li-Ping KONG Xue-Li ZHAO Xian-Jun HUA Jin-Lian 杜佳欣;李巴仑;寇正;吴礼平;孔学礼;赵献军;华进联(杨凌职业技术学院动物工程分院,杨陵712100;西北农林科技大学动物医学院/陕西省干细胞工程技术中心,杨陵712100)

机构地区：[1]Department of Animal Engineering,Yangling Vocational&Technical College,Yangling 712100,China [2]College of Veterinary Medicine,Shaanxi Centre of Stem Cells Engineering&Technology,Northwest A&F University,Yangling 712100,China

出　　处：《农业生物技术学报》2024年第12期2773-2785,共13页Journal of Agricultural Biotechnology

基　　金：杨凌职业技术学院科学研究基金项目(Zk23-15)。

摘　　要：Mesenchymal stem cells(MSCs)have the characteristics of promoting tissue repair,activating cytokine secretion,regulating organism immunity,and low moral and ethical constraints,which can provide effective treatment for various complex diseases in veterinary clinic.At present,MSCs are often cultured in a medium containing 10%fetal bovine serum(FBS),and the high cost of serum limits the mass production and application of MSCs.In this study,MSCs with a density of 70%were treated withα-MEM medium with gradient FBS of 0.5%~10%respectively.Cells were collected at 6,12,18,24 and 48 h for Giemsa staining,EDU(5 ethynyl-2'deoxyuridine)staining and cell migration experiments,respectively.The results showed that Serum Starvation(SS)of 2%FBS for 6 h could promote the migration ability of MSCs in vitro.Twelve diabetic dogs(Canine lupus familiaris)were given 1×10^(7) MSCs and SS-MSCs by intravenous injection,and the changes of blood sugar and weight of dogs were monitored within 2 weeks.Serological examination and histological examination were carried out 2 weeks after stopping treatment.The results showed that SS-MSCs group achieved similar therapeutic effects to normal culture environment,including restoring blood sugar and weight of diabetic dogs,remodeling islets,and improving liver glucose metabolism.This study provides a feasible strategy for clinical large-scale preparation of medical-grade MSCs,and improves the application value of MSCs in clinical disease treatment.间充质干细胞(mesenchymal stem cells,MSCs)具有促进组织修复、激活细胞因子分泌、调节机体免疫、道德伦理约束性低等特点,能为兽医临床上多种复杂病症提供有效治疗手段。目前,MSCs培养常采用含10%胎牛血清(fetal bovine serum,FBS)的培养液,高昂的血清成本限制了MSCs的批量生产与应用。本研究中,密度为70%的MSCs分别用含有0.5%~10%FBS的α-MEM培养基处理,分别在6、12、18、24和48 h收集细胞,进行Giemsa染色、5-乙炔基-2'脱氧尿苷(5 ethynyl-2'deoxyuridine,EDU)染色和细胞迁移实验。结果显示,2%FBS的血清饥饿(serum starvation,SS)处理6 h能促进MSCs的体外迁移能力。对12只糖尿病犬(Canis lupus familiaris)静脉移植1×10^(7) MSCs和SS-MSCs,监测犬的血糖与体重变化,治疗后2周进行血清学与组织学检查。结果显示,SS-MSCs组实现了与正常培养环境相似的治疗效果,包括恢复糖尿病犬的血糖和体重、重塑胰岛、提高肝脏葡萄糖代谢能力。本研究为临床大规模制备药用级MSCs提供了可行的策略,提高了MSCs在临床疾病治疗中的应用价值。

关 键 词：Serum starvation Mesenchymal stem cells Diabetes mellitus Canine lupus familiaris 

分 类 号：S856.5[农业科学—临床兽医学] Q28[农业科学—兽医学]