| 杜国庆,桑晓文,王 楠,石 瑛,熊轶喆,杨光月,陈元川,詹红生.大鼠半月板纤维软骨细胞的体外培养和生物学特征鉴定[J].现代生物医学进展英文版,2019,19(15):2829-2833. |
| 大鼠半月板纤维软骨细胞的体外培养和生物学特征鉴定 |
| Culture and Characterization of Rat Meniscus Fibrochondrocytes in Vitro |
| Received:November 24, 2018 Revised:December 18, 2018 |
| DOI：10.13241/j.cnki.pmb.2019.15.006 |
| 中文关键词: 半月板 纤维软骨细胞 体外培养 生物学鉴定 |
| 英文关键词: Meniscus Fibrochondrocytes Culture Characterization |
| 基金项目:国家自然科学基金项目青年科学基金项目（81704103）；"上海市重中之重临床重点学科建设项目"中医骨伤科学"（2017ZZ02024）；"工效筋骨学"上海市中医药新兴交叉学科资助计划建设项目（沪卫计中发2017【024】号）；上海市中西医临床协作试点项目"颈椎病"（ZXYXZ-201703） |
| Author Name | Affiliation | E-mail | | DU Guo-qing | 1 Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai, 201203, China
2 Institute of Traumatology & Orthopedics, Shanghai Academy of TCM, Shanghai, 201203, China | 530427540@qq.com | | SANG Xiao-wen | 1 Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai, 201203, China
2 Institute of Traumatology & Orthopedics, Shanghai Academy of TCM, Shanghai, 201203, China | | | WANG Nan | 1 Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai, 201203, China
2 Institute of Traumatology & Orthopedics, Shanghai Academy of TCM, Shanghai, 201203, China | | | SHI Ying | 1 Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai, 201203, China
2 Institute of Traumatology & Orthopedics, Shanghai Academy of TCM, Shanghai, 201203, China | | | XIONG Yi-zhe | 1 Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai, 201203, China
2 Institute of Traumatology & Orthopedics, Shanghai Academy of TCM, Shanghai, 201203, China | | | YANG Guang-yue | 1 Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai, 201203, China
2 Institute of Traumatology & Orthopedics, Shanghai Academy of TCM, Shanghai, 201203, China | | | CHEN Yuan-chuan | 1 Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai, 201203, China
2 Institute of Traumatology & Orthopedics, Shanghai Academy of TCM, Shanghai, 201203, China | | | ZHAN Hong-sheng | 1 Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai, 201203, China
2 Institute of Traumatology & Orthopedics, Shanghai Academy of TCM, Shanghai, 201203, China | |
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| 中文摘要: |
| 摘要 目的：体外分离、培养和鉴定大鼠半月板纤维软骨细胞, 为研究大鼠半月板纤维软骨细胞的损伤修复提供简单、可行的细胞培养方法。方法：机械分离大鼠膝关节内外侧半月板，0.1%Ⅱ型胶原酶消化配合机械吹打，10% FBS的培养基行原代和传代培养, 倒置显微镜动态观察不同时间点半月板纤维软骨细胞的形态及生长情况，鉴定采用甲苯胺蓝染色法和Ⅱ型胶原免疫细胞化学染色法。CCK-8法检测大鼠纤维软骨细胞增殖。结果：在不同时间点，细胞表现出不同的生理形态，由多角形或短梭形变为梭形，最终呈现出三角形或椭圆形，并随着时间延长，细胞增殖能力逐渐增加。细胞培养48 h和72 h的OD值分别明显高于培养24 h的OD值（P<0.05），差异具有统计学意义。细胞培养48 h和72 h的OD值相比较，72 h的OD值虽有增加，但并无统计学差异（P>0.05）。经甲苯胺蓝染色和Ⅱ型胶原免疫荧光染色为阳性。结论：该方法培养的细胞形态稳定，增殖能力强，具有体内纤维软骨细胞的生物学基本特性，可为后续实验提供简单、可靠的原代大鼠半月板纤维软骨细胞培养方法。 |
| 英文摘要: |
| ABSTRACT Objective: We want to provide a simple and feasible cell culture method for the future study of injury repair of rat meniscus by isolation, culture and identification of rat meniscus fibrochondrocytes in vitro. Methods: The lateral meniscus were mechanical separation and digestion by 0.1% Ⅱ collagenase, cultured in the 10% FBS medium. The morphology and growth situation of meniscal cells were observed by the inverted microscope dynamic at different time points, the identification of meniscus fibrochondrocytes by using toluidine blue staining and Ⅱ type collagen immune cell chemical dyeing method. The proliferation capacity of fibrochondrocytes was detected by CCK-8 method. Results: At different time points, the cells showed different physiological morphology, which was transformed from polygonal or short shuttle-shaped to fusiform, and finally appeared triangular or ellipsoidal, and the proliferation capacity of the cells increased gradually with the extension of time. The OD value at 48 h and 72 h in cell culture was significantly higher than that at 24 h (P<0.05), and the difference was statistically significant. The OD value of cell culture at 48 h and 72 h was increased, but there was no significant difference (P>0.05). By toluidine blue staining and collagen type Ⅱ immunofluorescence stain is positive. Conclusion: Cultured cells by this method are stable in morphology, strong in proliferation, and have the basic biological characteristics of fibrochondrocytes in vivo, which can provide a simple and reliable method for rat meniscus fibrochondrocytes culture. |
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