人咽缩肌细胞体外复合培养的研究  

In vitro Culture of Human Musculus Constrictor Pharyngis Superior Cells for Tissue Engineering Research

作　　者：陈静静[1] 沈志森[2] 竺亚斌[1] 康骋[1] 

机构地区：[1]宁波大学医学院,315211 [2]宁波市医疗中心李惠利医院,315040

出　　处：《医学研究杂志》2013年第3期85-89,共5页Journal of Medical Research

基　　金：浙江省杰出青年团队项目(R2101166);宁波市重大(重点)课题(2008C50019)

摘　　要：目的探索人咽缩肌细胞体外培养的条件,观察其在相应的生物支架上生长情况,为体外构建人下咽肌组织瓣做前期研究。方法取9例下咽癌志愿者癌旁正常的咽缩肌组织进行原代培养,当细胞传代至第4代时,将细胞接种于改性的1mm3平面或立体凹槽聚氨酯(polyurethane,PU)生物支架上进行复合培养。通过倒置相差显微镜、免疫荧光和电镜观察鉴定细胞的形态及增殖、分化的状况。结果咽缩肌最佳的体外培养条件是37℃、5%CO2孵箱环境,选择DMEM培养基并添加20%胎牛血清,采用组织块培养法可获得足够数目成活的咽缩肌细胞,此细胞在接枝丝素蛋白的凹槽PU支架上增殖分化良好并具备定向生长的能力,从而模拟了体内生长的状况。结论 组织块培养法获得的咽缩肌细胞与接枝丝素蛋白的凹槽PU支架复合培养时其生长状况佳,这为体外构建下咽组织瓣提供了一定的基础。Objective To explore in vitro culture condition of human pharyngeal constrictors cell, observe its growth status in the polymeric scaffolds. Methods Human pharyngeal constrictors cells from body tissue of nine volunteers of hypopharyngeal carcinoma pa- tients were isolated by explanting method. The 4th generation ceils for growthing on modified polyurethane （ PU ） scaffolds were collected with linearly aligned microgrooves. The cell morphology and the status of the proliferation and differentiation were identificated by morphol- ogy observation and immunohistochemistry analysis. Results Cells were cultured in Dulbecco＇s modified Eagle＇s medium （DMEM） , sup- plemented with 20% foetal bovine serum （FBS） under conditions of 37~C , 5% CO2 as the best condition for vitro culture. Silk fibroin （SF） grafting greatly improved scaffold＇s biological activity which led to the high cell biocompatibility. Conclusion The findings demon- strate that cells obtained from explanting method seeded on such scaffolds hold,which is promise for guiding human pharyngeal constrictors tissue construction.

关 键 词：人咽缩肌 复合培养 生物支架 丝素蛋白 

分 类 号：R-332[医药卫生]