中国医药
中國醫藥
중국의약
CHINA MEDICINE
2014年
10期
1522-1525
,共4页
软骨细胞%气管%原代细胞培养%体外生长
軟骨細胞%氣管%原代細胞培養%體外生長
연골세포%기관%원대세포배양%체외생장
Chondrocyte%Trachea%Primary cell culture%Growth pattern in vitro
目的 通过原代软骨细胞的体外培养和鉴定,探讨胰酶联合Ⅱ型胶原酶法体外分离培养软骨细胞的可行性.方法 分离出兔鼻中隔软骨组织,用胰酶联合Ⅱ型胶原酶的方法进行消化,获取原代软骨细胞.使用倒置显微镜观察软骨细胞的形态及生长情况,并用甲苯胺蓝染色、Ⅱ型胶原免疫组化及免疫荧光染色进行表型鉴定.结果 软骨细胞原代培养形成单层细胞需要7～8d,传代培养时间约2～3 d,细胞以圆形或类上皮细胞形态为主,甲苯胺蓝染色证实细胞可特异性合成糖胺聚糖,Ⅱ型胶原免疫组织化学染色及免疫荧光染色可证实细胞可特异性分泌Ⅱ型胶原.结论 本研究成功建立了简单易行的胰酶联合Ⅱ型胶原酶法体外分离培养大量纯净的软骨细胞,提高了消化速率,加大了细胞释放率,为组织工程气管软骨种子细胞的获取提供重要的技术支撑.
目的 通過原代軟骨細胞的體外培養和鑒定,探討胰酶聯閤Ⅱ型膠原酶法體外分離培養軟骨細胞的可行性.方法 分離齣兔鼻中隔軟骨組織,用胰酶聯閤Ⅱ型膠原酶的方法進行消化,穫取原代軟骨細胞.使用倒置顯微鏡觀察軟骨細胞的形態及生長情況,併用甲苯胺藍染色、Ⅱ型膠原免疫組化及免疫熒光染色進行錶型鑒定.結果 軟骨細胞原代培養形成單層細胞需要7～8d,傳代培養時間約2～3 d,細胞以圓形或類上皮細胞形態為主,甲苯胺藍染色證實細胞可特異性閤成糖胺聚糖,Ⅱ型膠原免疫組織化學染色及免疫熒光染色可證實細胞可特異性分泌Ⅱ型膠原.結論 本研究成功建立瞭簡單易行的胰酶聯閤Ⅱ型膠原酶法體外分離培養大量純淨的軟骨細胞,提高瞭消化速率,加大瞭細胞釋放率,為組織工程氣管軟骨種子細胞的穫取提供重要的技術支撐.
목적 통과원대연골세포적체외배양화감정,탐토이매연합Ⅱ형효원매법체외분리배양연골세포적가행성.방법 분리출토비중격연골조직,용이매연합Ⅱ형효원매적방법진행소화,획취원대연골세포.사용도치현미경관찰연골세포적형태급생장정황,병용갑분알람염색、Ⅱ형효원면역조화급면역형광염색진행표형감정.결과 연골세포원대배양형성단층세포수요7～8d,전대배양시간약2～3 d,세포이원형혹류상피세포형태위주,갑분알람염색증실세포가특이성합성당알취당,Ⅱ형효원면역조직화학염색급면역형광염색가증실세포가특이성분비Ⅱ형효원.결론 본연구성공건립료간단역행적이매연합Ⅱ형효원매법체외분리배양대량순정적연골세포,제고료소화속솔,가대료세포석방솔,위조직공정기관연골충자세포적획취제공중요적기술지탱.
Objective To explore the feasibility of seeding cells in tissue engineering trachea through primary culture and identification of cartilage cells in vitro.Methods The primary cartilage cells were isolated from the choelroseptum of rabbit,and digested by using trypsogen associated with type-Ⅱ collagenase.The morphology and growth of cartilage cells were observed by inverted microscope,and identificated by the use of toluidine blue staining,the immunohistochemistry and immunofluorescence of type-Ⅱ collagen.Results The time of form monolayer cells was 7-8 days and subculture was 2-3 days.The morphology of cartilage cells mainly consisted of circular and epithelioid cells.The cell-specific glycosaminoglycan was confirmed by the use of the method of the toluidine blue staining.The cell-specific secrete type-Ⅱ collagen was confirmed by using the immunohistochemistry and the immunofluorescence of type-Ⅱ collagen.Conclusions This study successfully establishes a simple method for isolating and cultivating chondrocytes by using the trypsinogen and the type-Ⅱ collagenase.This method may improve the digestion rate and increase the cells release rate to provide the important technical support for tissue engineered trachea seed cells.
