中国骨质疏松杂志
中國骨質疏鬆雜誌
중국골질소송잡지
CHINESE JOURNAL OF OSTEOPOROSIS
2015年
4期
409-414
,共6页
许伟%谢兴文%赵永利%李宁%徐世红%宋敏
許偉%謝興文%趙永利%李寧%徐世紅%宋敏
허위%사흥문%조영리%리저%서세홍%송민
Brdu%rBMSCs%免疫荧光%迁移
Brdu%rBMSCs%免疫熒光%遷移
Brdu%rBMSCs%면역형광%천이
BrdU%rBMSCs%Immunohistochemistry%Migration
目的:5-溴-2-脱氧尿嘧啶核苷(5-bromo-2-deoxyuridine,Brdu)体外标记大鼠骨髓间充质干细胞(rBMSCs),通过尾静脉回植于大鼠体内,观察其是否在大鼠体内能向损伤部位迁移。方法采用贴壁筛选法培养rBMSCs,对rBMSCs进行鉴定,用Brdu标记细胞并计算标记率;大鼠颅骨骨缺损模型的建立;Brdu标记的rBMSCs体内回植;分别于7、14、21d后处死大鼠取出损伤部位的颅骨,并对其脱钙、制作切片;做荧光免疫组化染色;用荧光倒置相差显微镜来观察标本处是否有Brdu标记的rBMSCs。结果筛选纯化的骨髓间充质干细胞呈均一的成纤维细胞样,贴壁生长,以长梭形为主,漩涡状盘旋排列,CD45阴性表达,CD44、CD90表达阳性,rBMSCs经诱导后可向成脂、成骨分化。 Brdu标记率为86.2%,在标本处检测到Brdu标记的rBMSCs,14d迁移效果最佳。结论 Brdu标记的rBMSCs可以进行大鼠体内的迁移研究。
目的:5-溴-2-脫氧尿嘧啶覈苷(5-bromo-2-deoxyuridine,Brdu)體外標記大鼠骨髓間充質榦細胞(rBMSCs),通過尾靜脈迴植于大鼠體內,觀察其是否在大鼠體內能嚮損傷部位遷移。方法採用貼壁篩選法培養rBMSCs,對rBMSCs進行鑒定,用Brdu標記細胞併計算標記率;大鼠顱骨骨缺損模型的建立;Brdu標記的rBMSCs體內迴植;分彆于7、14、21d後處死大鼠取齣損傷部位的顱骨,併對其脫鈣、製作切片;做熒光免疫組化染色;用熒光倒置相差顯微鏡來觀察標本處是否有Brdu標記的rBMSCs。結果篩選純化的骨髓間充質榦細胞呈均一的成纖維細胞樣,貼壁生長,以長梭形為主,漩渦狀盤鏇排列,CD45陰性錶達,CD44、CD90錶達暘性,rBMSCs經誘導後可嚮成脂、成骨分化。 Brdu標記率為86.2%,在標本處檢測到Brdu標記的rBMSCs,14d遷移效果最佳。結論 Brdu標記的rBMSCs可以進行大鼠體內的遷移研究。
목적:5-추-2-탈양뇨밀정핵감(5-bromo-2-deoxyuridine,Brdu)체외표기대서골수간충질간세포(rBMSCs),통과미정맥회식우대서체내,관찰기시부재대서체내능향손상부위천이。방법채용첩벽사선법배양rBMSCs,대rBMSCs진행감정,용Brdu표기세포병계산표기솔;대서로골골결손모형적건립;Brdu표기적rBMSCs체내회식;분별우7、14、21d후처사대서취출손상부위적로골,병대기탈개、제작절편;주형광면역조화염색;용형광도치상차현미경래관찰표본처시부유Brdu표기적rBMSCs。결과사선순화적골수간충질간세포정균일적성섬유세포양,첩벽생장,이장사형위주,선와상반선배렬,CD45음성표체,CD44、CD90표체양성,rBMSCs경유도후가향성지、성골분화。 Brdu표기솔위86.2%,재표본처검측도Brdu표기적rBMSCs,14d천이효과최가。결론 Brdu표기적rBMSCs가이진행대서체내적천이연구。
Objective Rat bone marrow mesenchymal stem cells ( rBMSCs) were labeled with 5-bromo-2-deoxyuridine ( BrdU) in vitro and were transplanted in rats from the tail vein.The cell migration to the damage part was observed.Methods rBMSCs were cultured using adherent screening method.Phenotypic characterization of rBMSCs was identified.The cells were labeled with BrdU and the rate of labeling was calculated.The rat skull defect model was established.The BrdU-labeled rBMSCs were transplanted into the rats.The rats were sacrificed on 7, 14, and 21 days, respectively, and the defect skull was collected, decalcified, and sliced.BrdU-labeled rBMSCs were observed using immunohistochemistry.Results Screened rBMSCs appeared as unique fibroblastic-like cells.Cells were adherent to the wall, with long spindle shape and swirling arrangement.The expression of CD45 was negative, but the expressions of CD44 and CD90 were positive.The rBMSCs had the ability of differentiation to adipocytes and osteoblasts.The rate of BrdU labeling was 86.2%.The BrdU labeling was detected at the defects.The migration was the best at the 14 days.Conclusion The BrdU-labeled rBMSCs can be used in rat cell migration study.