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山东大学学报(医学版) ›› 2012, Vol. 50 ›› Issue (12): 98-.

• 临床医学 • 上一篇    下一篇

微载体旋转立体培养对成人退变髓核细胞凋亡调控的研究

宁斌1, 龚维明1, 李玲梅2, 郭洁1, 夏启杆1, 宁红1, 贾堂宏1   

  1. 1.山东大学附属济南市中心医院脊柱外科, 济南 250013; 2.济南市急救中心,济南 250013
  • 收稿日期:2012-03-29 出版日期:2012-12-10 发布日期:2012-12-10
  • 通讯作者: 贾堂宏(1957- ),男,主任医师,教授,博士生导师,主要从事脊柱退变的研究。E-mail: jiatanghong@hotmail.com
  • 作者简介:宁斌(1979- ),男,主治医师,博士研究生,主要从事脊柱疾患的研究。E-mail: ningbin@mail.sdu.edu.cn

Effect of stiring microcarrier system culturing on apoptosis regulation
of adult degenerative nucleus pulposus cells

NING Bin1, GONG Wei-ming1, LI Ling-mei2, GUO Jie1, XIA Qi-gan1, NING Hong1, JIA Tang-hong1   

  1. 1. Department of Spinal Surgery, Jinan Central Hospital Affiliatted to Shandong Universtiy, Jinan 250013, China;
    2. The Emergency Center of Jinan, Jinan 250013, China
  • Received:2012-03-29 Online:2012-12-10 Published:2012-12-10

摘要:

目的   采用微载体旋转立体培养法对成人退变髓核细胞进行扩增,研究细胞凋亡情况,探讨髓核细胞凋亡在椎间盘退变中的作用。方法   手术切除的退变髓核组织行原代培养,传代后随机进行单层培养和微载体旋转立体培养,采用Annexin-V-PI染色,流式细胞仪定量检测早期细胞凋亡,应用免疫组化方法检测凋亡抑制因子Bcl2的表达,DeadEndTM比色法检测细胞晚期凋亡情况。结果   流式细胞仪定量检测可见细胞在稳定生长期时,微载体旋转立体培养法较单层培养法髓核细胞的凋亡率低(P<0.05),在对数生长期两种培养方法无明显差别(P>0.05)。Bcl-2指标及DeathEndTM比色法检测显示在两个时期内,微载体旋转立体培养法培养的细胞凋亡率较低(P<0.05)。结论   微载体旋转立体培养法可较好地降低髓核细胞凋亡率,是一种较有前途的髓核细胞培养方法,为组织工程化椎间盘髓核种子细胞的研究奠定了基础。

关键词: 微载体;细胞凋亡;细胞培养;髓核;退变

Abstract:

Objective   To investigate the effects of nucleus pulposus cells apoptosis on degenerated intervertebral discs by using cell amplification with microcarrier system and studying cell apoptosis. Methods   After primary culture of the degenerative nucleus pulposus tissues by operation, nucleus pulposus cells were randomly separated into two groups. Of the two groups, one was cultured with monolayer cluture, another with microcarrier system. In order to detect apoptosis of different stages, Annexin-V-PI staining was used for early stage apoptosis, DeadEndTM staining for late stage apoptosis, immunohistochemical staining for Bcl2 protein. Results   Annexin-V-PI staining demonstrated that apoptosis rate of nucleus pulposus cultured with microcarrier system was lower in stable phase (P<0.05). However, there was no significant difference between the two different culture methods in logarithmic growth phase (P>0.05).  The data of DeadEndTM staining and immunohistochemical staining showed that apoptosis rate of nucleus pulposus cultured with microcarrier system was lower in both phases (P<0.05). Conclusion   Microcarrier system, which can efficiently reduce the apoptosis rate of nucleus pulposus cells, is a promising culture method applied in tissue engineering of intervertebral discs.

Key words: Microcarrier; Apoptosis; Cell culture; Neucleus pulposus; Degenerative

中图分类号: 

  • R681.55
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