基础医学与临床 ›› 2014, Vol. 34 ›› Issue (12): 1682-1688.

• 研究论文 • 上一篇    下一篇

CDMP1转基因间充质干细胞片修复兔软骨缺损

崔颖1,姚梅1,胡晶1,李俊霞1,王宇1,张本2   

  1. 1. 辽宁医学院附属第一医院
    2.
  • 收稿日期:2014-03-27 修回日期:2014-07-01 出版日期:2014-12-05 发布日期:2014-11-25
  • 通讯作者: 崔颖 E-mail:yingwu2002@163.com
  • 基金资助:
    转基因细胞片修复软骨缺损的研究;TAT-PDX1蛋白诱导的骨髓间充质干细胞分化为胰岛样细胞的研究

Repairing larynx cartilage defects with MSCs cell sheets transfected by CDMP1

  • Received:2014-03-27 Revised:2014-07-01 Online:2014-12-05 Published:2014-11-25
  • Contact: CUI Ying E-mail:yingwu2002@163.com

摘要: 目的 探讨软骨源性形态发生蛋白-1(CDMP-1)转基因细胞片修复兔软骨缺损的能力。方法 腺病毒转染CDMP1基因至第4代兔骨髓间充质干细胞(BMSCs)中,连续培养7~14d后接种至温度敏感性培养皿中制备细胞片,Real time PCR和Western blot法检测转基因细胞片的CDMP1及Π型胶原蛋白表达;用细胞片工程技术构建组织工程化细胞片并移植入兔甲状软骨缺损处修复软骨缺损。实验分(A)BMSCs细胞片组;(B)转染Ad-CMV-eGFP细胞片组;(C)转染Ad-CMV-hCDMP1-IRES-eGFP细胞片组。分别于术后4和8周检测工程化软骨的Π型胶原蛋白及蛋白多糖(GAG)表达。结果检测到转基因细胞片中CDMP1的表达;所获得的工程化软骨免疫组化和阿利新蓝染色显示: A组Π型胶原蛋白和GAG表达阳性, B组和C组Π型胶原蛋白和GAG表达阴性(P<0.05)。结论 CDMP转基因细胞片具有较好的软骨分化活性,可有效地修复兔喉软骨缺损。

关键词: 细胞片, CDMP1, BMSCs, 腺病毒转染, 软骨组织工程

Abstract: Objective To detect the repair capacity in rabbit cartilage defects by BMSCs cell sheets transfected by CDMP. Methods Rabbit bone marrow mesenchymal stem cells were Isolated and cultured in vitro; the passage 4 cells were tansfected with exogenous recombinant human cartilage-derived morphogenetic protein 1 by adenovirus vector; the transgenosis cell sheets were harvested by means of temperature-responsive culture dish after 7-14 days. After detecting the express of CDMP1 in transgenic cell sheet using Real Time PCR and Western blot, then the sheets were implanted into the rabbit thyroid cartilage defects .After 4 or 8 weeks, the culturing systems were analyzed at the gross level as well as at the histological level to observe the repair ability.The experiment was divide into three groups: (A)transfected by Ad-CMV-hCDMP1-IRES-eGFP BMSCs cell sheets;(B)transfected by Ad-CMV -eGFP BMSCs cell sheets;(C)BMSCs cell sheets . Results The express of CDMP1 in transgenosis cell sheets can be detected by Real Time PCR and Western blot; The detection of immunohistochemistry and Alcian Blue staining showed the expression of collagen II and GAG in group A,while negative expression in group B and group C. There was statistical difference between group A, group B and C (P<0.05). Conclusion CDMP transgenosis cell sheets had superior chondrogenic capacity ,which can effectively repair rabbit thyroid cartilage defects.

Key words: Cell sheet, CDMP1, BMSCs, Adenovirus infection, Cartilage tissue engineering

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