分离扩增肋软骨细胞在三维支架上的种植与培养  

Implantation and culture of rib chondrocytes on a three-dimensional scaffold

作　　者：刘政呈[1] 郭翔[1] 赵珩[1] 

机构地区：[1]上海市胸科医院胸外科,上海市200030

出　　处：《中国组织工程研究与临床康复》2010年第34期6297-6300,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：上海市胸科医院胸外科重点课题~~

摘　　要：背景:外科长段气管切除后,需要应用气管替代物,理想的替代材料需具有相当的强度、可弯曲性以及内面覆盖的上皮。合适的种子细胞、性质优良的载体、良好的生长媒介是组织工程化气管构建中最重要的组成部分。目的:寻找分离培养肋软骨细胞的最佳方法,构建软骨-支架模型。方法:将取出的兔第5,6肋软骨切成组织片,经胰酶、蛋白酶消化或酶处理后贴壁,分离肋软骨细胞并扩增。传代2次后种植于聚乳酸羟基乙酸或涤纶支架上,继续载体外环境下培养。观察单层培养扩增中软骨细胞的形态、结构,及在支架上培养时的细胞生长状态、外分泌基质。结果与结论:酶消化法和组织块培养法所获细胞均为原代肋软骨细胞,单次传代时间为5d,但组织块培养法分离细胞较慢,且存在成纤维细胞污染情况,提示酶消化法适合用于肋软骨细胞的分离。肋软骨细胞种植于支架后贴附良好,第2周时可见基质分泌,获得软骨-支架模型,可用于构建组织工程化气管。BACKGROUND：Long-segment tracheal resection surgery requires the application of tracheal replacement.The ideal substitute materials should be present with considerable strength,flexibility and inner surface-covered epithelium.Suitable seed cells,excellent carrier,good growth medium are the key components of tissue-engineered trachea construction.OBJECTIVE：To investigate optimal way for rib chondrocytes isolation and culture,and to construct cartilage-scaffold models.METHODS：Rabbits＇ 5th and 6th rib cartilages were sliced into 1-mm3 pieces and treated with sequential enzyme digest or block culture to isolate chondrocytes.After two passages,rib chondrocytes were collected and suspended.The cell suspension was seeded on poly（lactic acid-glycolic acid） or Dacron scaffold,then cultured under a 37 ℃,5% CO2 atmosphere.Chondrocytes＇ morphology and structure during monolayer culture expansion were observed,while cells growing and matrices formation on scaffold were examined.RESULTS AND CONCLUSION：Cells isolated by sequential enzyme digest or block culture were primary rib chondrocytes,of which the average passage time was 5 days.However,isolation by tissue block culture was slow and even fibroblasts were found in one flask,indicating that enzyme digest fits for the rib chondrocyte isolation.Chondrocytes attached well on scaffolds.Extracellular matrices were seen to be secreted after 2 weeks.The chondrocytes-scaffold model could be further used to construct tissue-engineered trachea.

关 键 词：软骨 培养 支架 酶消化法 生物材料 组织块培养法 组织工程 

分 类 号：R318[医药卫生—生物医学工程]