中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (45): 8434-8438.doi: 10.3969/j.issn.1673-8225.2011.45.016

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

预损伤与改良传代体外培养许旺细胞的比较

杨  俊1，秦建强2，裴国献3，郭新辉1，叶汉深1，陈大军1，洪加津1，蔡伟斌1   

  1. 1解放军第四二一医院骨科，广东省广州市  510310
    2南方医科大学临床解剖学研究所广东省组织构建与检测重点实验室，广东省广州市  510515
    3解放军第四军医大学附属西京骨科医院，陕西省西安市  710032
  • 收稿日期:2011-08-02 修回日期:2011-09-07 出版日期:2011-11-05 发布日期:2011-11-05
  • 通讯作者: 秦建强，博士，教授，南方医科大学临床解剖学研究所广东省组织构建与检测重点实验室，广东省广州市 510515
  • 作者简介:杨俊★，男，1977年生，辽宁省辽阳市人，汉族，2005年解放军第一军医大学毕业，硕士，主治医师，主要从事创伤骨科、脊柱外科的临床诊治、周围神经损伤及军事训练伤的科学研究。

Comparison of pre-injury and improved passage methods for Schwann cells cultured in vitro

Yang Jun1, Qin Jian-qiang2, Pei Guo-xian3, Guo Xin-hui1, Ye Han-shen1, Chen Da-jun1, Hong Jia-jin1, Cai Wei-bin1   

  1. 1Department of Orthopedics, the 421 Hospital of Chinese PLA, Guangzhou  510310, Guangdong Province, China
    2Institute of Clinical Anatomy, Southern Medical University, Guangdong Provincial Key Laboratory of Tissue Construction and Inspection, Guangzhou  510515, Guangdong Province, China
    3Xijing Hospital, the Fourth Military Medical University of Chinese PLA, Xi'an  710032, Shaanxi Province, China
  • Received:2011-08-02 Revised:2011-09-07 Online:2011-11-05 Published:2011-11-05
  • Contact: Qin Jian-qiang, Ph.D., Professor, Institute of Clinical Anatomy, Southern Medical University, Guangdong Provincial Key Laboratory of Tissue Construction and Inspection, Guangzhou 510515, Guangdong Province, China jqqin@fimmu.com
  • About author:Yang Jun★, Master, Attending physician, Department of Orthopedics, the 421 Hospital of Chinese PLA, Guangzhou 510310, Guangdong Province, China yangjun1267@sina.com

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摘要：

背景：通过组织工程方法构建神经桥接体修复神经损伤，需要大量纯化体外培养的许旺细胞。
目的：对比观察预损伤法和改良传代法获取许旺细胞的纯度与质量。
方法：①预损伤法：预损伤SD乳鼠坐骨神经，3 d后取出坐骨神经，分离神经外膜，用胰酶、胶原酶消化，差速贴壁除去成纤维细胞，接种培养。②改良传代法：直接获取SD乳鼠坐骨神经，分离神经外膜，运用双酶消化法结合单酶消化法进行许旺细胞原代培养，5~7 d后采用单酶快速消化离心法行传代培养，同时纯化许旺细胞。
结果与结论：预损伤法和改良传代法体外培养的许旺细胞纯度均达95%以上，两种方法获得的许旺细胞纯度差异无显著性意义(P > 0.05)。两种方法获取的许旺细胞形态正常，数量及纯度高，增殖旺盛，说明预损伤法和改良传代法都是体外获取高质量与高纯度许旺细胞的理想方法。

关键词: 许旺细胞, 细胞培养, 组织工程, 预损伤法, 改良传代法 

Abstract:

BACKGROUND: Tissue engineering methods to construct nerve bridge graft for repairing nerve injury requires massive purified Schwann cells cultured in vitro.
OBJECTIVE: To compare the purity and quality of Schwann cells obtained with the pre-injury method and improved passage method.
METHODS: (1) Pre-injury method: Sciatic nerve of Sprague-Dawley neonatal rats was pre-injured for 3 days and removed, the epineurium was isolated and digested with trypsin and collagenase. Schwann cells were cultured in the culture medium after fibroblasts were eliminated with differential adhesion method. (2) Improved passage method: Sciatic nerve was directly removed from Sprague-Dawley neonatal rats and the epineurium was isolated. Double-enzyme digestion and single-enzyme digestion methods were adopted to primarily culture Schwann cells for 5-7 days. Schwann cells were purified with a single enzyme rapid digestion and centrifugation.
RESULTS AND CONCLUSION: Both pre-injury and improved passage methods obtained 95% purity of Schwann cells cultured in vitro, with no significant difference (P > 0.05). The resultant Schwann cells obtained by two methods showed normal morphology, high quantity, high purity and productive growth. These experimental findings imply that both pre-injury and improved passage methods are ideal approaches for in vitro culture of Schwann cells with high quality and high purity.

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