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| 探讨年龄因素对人黄韧带细胞原代培养的影响 |
| 张宇1,吴少鹏1,夏雄智1,邓崇礼1,李参天1,许学猛1,钟招明2,陈建庭2* |
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(1.广东省第二中医院脊柱骨科, 广州 510095
2.南方医科大学南方医院脊柱骨病外科,广州 510515
*通信作者) |
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| 摘要: |
| 目的 探讨从不同年龄的人获得黄韧带组织与原代培养细胞游出之间的关系, 以提高黄韧带细胞原代培养的成功率。方法 采用酶消化加组织块培养法,对来自21例不同年龄腰椎管狭窄和腰椎间盘突出症患者的黄韧带组织进行黄韧带细胞原代培养, 倒置相差显微镜下观察细胞从组织块迁出时间、形态和生长状态；采用细胞免疫荧光染色法对传第3代细胞进行Ⅰ型胶原和波形蛋白表达鉴定；对组织块内细胞游出率和年龄进行相关性分析。结果 倒置显微镜下见黄韧带细胞形态生长良好,培养细胞的波形蛋白和Ⅰ型胶原免疫荧光化学染色均呈阳性。患者年龄与黄韧带组织块原代培养时细胞游出率之间呈负相关关系(r=-0.618, P<0.05),年轻患者黄韧带组织细胞游出率高。结论 原代培养人黄韧带细胞时尽量采用年轻人黄韧带组织。 |
| 关键词: 黄韧带 细胞培养技术 年龄因素 组织块加酶消化法 |
| DOI：10.3724/SP.J.1008.2012.099 |
| 投稿时间：2011-04-18修订日期：2011-11-04 |
| 基金项目:广东省自然科学基金博士启动项目(S2011040005276),广东省中医药局建设中医药强省立项资助课题(20111124). |
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| Effect of age on primary culture of human ligamentum flavum cells |
| ZHANG Yu1,WU Shao-peng1,XIA Xiong-zhi1,DENG Chong-li1,LI Can-tian1,XU Xue-meng1,ZHONG Zhao-ming2,CHEN Jian-ting2* |
(1. Department of Orthopaedic and Spinal Surgery, Second Provincial Traditional Chinese Medicine Hospital of Guangdong, Guangzhou 510095, Guangdong, China
2. Department of Orthopaedic and Spinal Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong, China
*Corresponding author.) |
| Abstract: |
| Objective To study the relationship of different ages with harvesting of ligamentum flavum and cell growth rate in primary culture, so as to improve the success rate of human ligamentum flavum cells(LFCs) culture in vitro. Methods A total of 21 surgical ligamentum flavum specimens from patients with intervertebral disc herniation and lumbar spinal stenosis were primary cultured with tissue block enzymolytic method. Cell outgrowth, morphology and growth status were observed under an inverted phase microscope. Expression of vimentin and type Ⅰ collagen was detected in the cells of third passage by immunofluorescence staining. The correlation between age and cell growth rate was analyzed. Results Cell morphology and growth status were well under inverted phase microscope and immunofluorescence staining showed vimentin and collagen type Ⅰ expression in the all LFCs. The cell growth rate of primary culture was negatively correlated with the age of patients from whom ligamentum flavum had been collected(r=-0.618, P<0.05). Cell growth rate was high when the patients were young. Conclusion Ligamentun flavum specimens should be taken from young subjects to ensure the success rate of primary culture of LFCs. |
| Key words: ligamentum flavum cell culture techniques age factors tissue block enzymolytic method |
