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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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brdu体外标记大鼠骨髓间充质干细胞的研究
, PP. 184-186
Keywords: 尿嘧啶核苷酸类,流式细胞术,细胞,培养的,细胞分离,造血干细胞,骨髓细胞
Abstract:
目的探讨连续培养的骨髓间充质干细胞(mscs)的5-溴脱氧尿嘧啶核苷(brdu)最佳标记时间、剂量。方法差速贴壁法对sd鼠mscs进行体外传代培养;第6代细胞行流式细胞仪鉴定细胞的表面抗原;以终浓度分别为5、10、15μmol/l的brdu检测最佳标记剂量;在细胞生长融合前72、48、36、24、12、3h加入brdu,使终浓度为10μmol/l,并分别孵育至细胞融合,测定brdu的标记率,找出最佳标记时间;免疫组化分析brdu标记率。结果流式细胞仪检测所标记的细胞表达cd29和cd44,不表达cd11b和cd45;终浓度为10μmol/l和孵育48hbrdu标记率均>98%,并且连续传5代均可检测到。结论传代后贴壁生长的梭形细胞为mscs,终浓度为10μmol/l和孵育48hbrdu标记率均>98%,且可以连续检测到,提示brdu标记可用于mscs移植入体内后存活、生长和分化的动态观察。
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