摘要
背景:骨髓间充质干细胞系统性输注后,何种因素促使其迁移到正确部位尤为关键,目前认为黏附分子在介导骨髓间充质干细胞向缺血或损伤组织迁移过程中起重要作用。目的:观察血管细胞黏附分子1与细胞间黏附分子1在大鼠骨髓间充质干细胞中的表达。方法:采用直接贴壁法体外分离培养大鼠骨髓间充质干细胞,免疫细胞化学染色检测血管细胞黏附分子1及细胞间黏附分子1蛋白的表达,应用免疫荧光直标法在流式细胞仪上检测血管细胞黏附分子1及细胞间黏附分子1抗原的表达率,RT-PCR半定量分析血管细胞黏附分子1及细胞间黏附分子1mRNA的表达。结果与结论:免疫细胞化学染色结果显示,骨髓间充质干细胞血管细胞黏附分子1呈弱阳性表达,细胞间黏附分子1呈强阳性表达。流式细胞仪检测结果显示,血管细胞黏附分子1表达率为6%,细胞间黏附分子1表达率为100%。RT-PCR检测结果显示,血管细胞黏附分子1mRNA呈微弱表达,细胞间黏附分子1mRNA呈高度表达。提示在生理状态下,体外培养的大鼠骨髓间充质干细胞低表达血管细胞黏附分子1,高表达细胞间黏附分子1。
BACKGROUND：Schwann cells are the seed cells of neural repair,and it is a key to harvest a large number of Schwann cells with high purity and activity.OBJECTIVE：To compare the in vitro culture,purification,and morphology of Schwann cells between neonatal and adult rats,and investigate a simple and feasible culture method to harvest high-purity Schwann cells.METHODS：Totally 30 Sprague-Dawley rats,comprising 20 neonatal （1-3 days after birth,neonatal group） and 10 adult （weighing 150-200 g,adult group） rats,were included.Following double-enzyme digestion and two incubations,Schwann cells were isolated and purified by differential attachment.Cell morphology and attaching speed were determined through the use of inverted microscope.Cells were counted and cell purity was calculated.Cell proliferative ability was detected by MTT microcolorimetry.Curves of cell proliferation in each group were depicted to determine proliferative speed.Schwann cells were identified by S-100 immunochemistry.RESULTS AND CONCLUSION：Compared with fibroblasts,neonatal rat Schwann cells exhibited faster,while adult rat Schwann cells showed slower,attaching speed.Both neonatal and adult groups yielded over 96% cell purity.MTT microcolorimetry results revealed that Schwann cells proliferated actively in neonatal and adult groups.Cell proliferative curves show that neonatal rat Schwann cells proliferated faster than adult rat Schwann cells （P 0.05）.S-100 immunochemistry results showed positive results in both groups.All these findings suggest that double-enzyme digestion and two incubations followed by differential attachment is a satisfactory method to harvest considerable Schwann cells with high purity and activity.Neonatal rat Schwann cells show stronger proliferative,attaching capacities than adult rat Schwann cells.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第6期1130-1133,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
