动态种植法提高在去细胞猪瓣支架上种植内皮细胞效率和均匀性的研究  

Study on improving endothelial cells seeding efficiency and uniformity on acellular porcine aortic valve scaffolds with oscillating seeding

作　　者：顾春虎[1] 王云雅[2] 易定华[1] 魏旭峰[1] 董小超[1] 刘洋[1] 金艳[1] 孙国成[1] 

机构地区：[1]第四军医大学西京医院心脏外科,陕西西安710033 [2]第四军医大学西京医院核医学科,陕西西安710033

出　　处：《第四军医大学学报》2008年第11期971-974,共4页Journal of the Fourth Military Medical University

基　　金：陕西省卫生厅基金(06049);国家自然科学基金(30600137;30672086);国家863计划(2006AA02A138)

摘　　要：目的:尝试使用简单的动态种植方法来提高内皮细胞在去细胞牛心包支架上的种植效率和均匀性,以更有效地构建组织工程瓣膜.方法:采用胰酶-去垢剂法制备去细胞牛心包.内皮细胞以下列方式在去细胞牛心包支架进行种植和培养:①振荡种植振荡培养(OO);②振荡种植静止培养(OS);③静止种植静止培养(SS).定期取样品进行电镜、组织学检测及细胞计数分析.结果:细胞加入24h后振荡种植的样品比静止种植的样品细胞在支架内分布更为均匀,同时振荡种植的样品细胞贴壁率[(36±3)%]高于静止种植的样品细胞贴壁率[(21±2)%,P<0.05].振荡种植振荡培养的样品在培养过程中细胞脱离支架而死亡,因此放弃了这些样品的继续培养.振荡种植静止培养的样品和静止种植静止培养的样品细胞一直保持正常的伸展状态.在培养过程中支架上的活细胞总数振荡种植静止培养的样品一直高于静止种植静止培养的样品.结论:采取振荡种植的方式种植细胞可明显提高内皮细胞在三维支架上种植的效率和分布均匀性.AIM： To explore a simple way to improve seeding efficiency and uniformity on acellular porcine aortic valve, in order to effectively construct tissue-engineered valve. METHODS： Cells of porcine aortic valves were removed by a trypsin-deterging cell extraction process. Endothelial ceils of thoracic aorta from Sprague-Dawley （SD） rat were added onto acellular scaffolds and cultured in the following 3 ways： oscillating seeding and oscillating culture （ OO）, oscillating seeding and static culture （ OS）, and static seeding and static culture （SS）. Samples were obtained and subjected to light microscopy, scanning electron microscopy （SEM） and quantitative cell number assay at planed times. RESULTS： Oscillating seeding achieved more uniform cell distribution within scaffolds, compared with static seeding. Moreover, Endothelial ceils seeding efficiency was significantly higher in oscillating conditions [ （ 36 ± 3 ） % ] than in static conditions [ （21 ± 2）%, P 〈0.01 ]. Ceils from OO group samples died due to their breaking away from the vascular frame and were discarded. Ceils from OS group samples and SS group samples remained a normal extension. The living cell number was higher in OS samples than that in SS throughout the culture. CONCLUSION： Compared with conventional static techniques, oscillating seeding of endothelial ceils on acellular scaffolds can obviously improve cell seeding efficiency and uniformity.

关 键 词：内皮细胞 振荡种植 静止种植 振荡培养 静止培养 

分 类 号：R318[医药卫生—生物医学工程]