一种可提高成纤维细胞在三维支架上种植效率和均匀性的简单方法  

A simple way to improve fibroblast seeding efficiency and uniformity in constructing 3-D dermal equivalent

作　　者：周娟[1] 张其清[1] 

机构地区：[1]中国医学科学院/协和医科大学生物医学工程研究所,天津300192

出　　处：《第二军医大学学报》2006年第1期101-104,共4页Academic Journal of Second Military Medical University

基　　金：国家高技术研究发展计划/863计划(2002AA326040).~~

摘　　要：目的:尝试使用简单的动态种植方法来提高成纤维细胞在三维支架上的种植效率和均匀性,以更有效地构建组织工程皮肤。方法:成纤维细胞以下列方式在胶原海绵支架进行种植和培养:(1)振荡种植振荡培养;(2)振荡种植静止培养;(3)静止种植静止培养。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24 h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[(56±5)%]明显高于静止种植的样品细胞贴壁率[(31±4)%,P<0.01]。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。在培养过程中细胞向支架内部的长入速度及支架中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成纤维细胞在三维支架上种植的效率和分布均匀性。Objective：To explore a simple way to improve seeding efficiency and uniformity in constructing 3-D dermal equivalent, in order to effectively construct tissue engineered skin. Methods： Human dermal fibroblasts were added onto collagen sponge scaffolds and cultured in the following 3 ways： （1）oscillating seeding and oscillating culture （OO）, （2）oscillating seeding and static culture （OS）, and （3）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 spatial cell distribution within scaffolds as compared to static seeding. Moreover, fibroblast seeding efficiency was significantly higher in oscillating conditions （[56±4 5]% ） than in static conditions（[31±4]%, P〈0.01 ）. Cells from OO samples died due to their breaking away from the dermal frame and were discarded. Cells from OS samples ahd SS samples remained a normal extension. The cell ingrowth and the cell number were both higher for OS samples than that for SS throughout the culture. Conclusion： These results suggest that, as compared to conventional static techniques, oscillating seeding of dermal fibroblasts on 3-D scaffolds can obviously improve seeding efficiency and uniformity.

关 键 词：成纤维细胞 振荡种植 静止种植 振荡培养 静止培养 

分 类 号：Q813.11[生物学—生物工程]