中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (33): 5339-5347.doi: 10.3969/j.issn.2095-4344.2017.33.015

• 干细胞移植 stem cell transplantation • 上一篇    下一篇

诱导多能干细胞对骨关节炎软骨损伤的修复作用

吴张松1,朱红霞2,罗志强1,吴晓敏1,林光耀1,张明裕1,顾洪生2,朱艳霞1   

  1. 1深圳大学医学部,深圳抗衰老与再生医学重点实验室,广东省深圳市 518060;2孝感市妇幼保健院脊柱外科,广东省深圳市 518060
  • 修回日期:2017-06-07 出版日期:2017-11-28 发布日期:2017-12-01
  • 通讯作者: 朱艳霞,博士,副教授,深圳大学医学部,深圳抗衰老与再生医学重点实验室,广东省深圳市 518060
  • 作者简介:吴张松,男,1995年生,安徽省宿松县人,汉族,深圳大学临床医学本科在读,主要从事干细胞与软骨组织工程研究。
  • 基金资助:

    国家自然科学基金(81301597);深圳新兴战略产业发展专项资金(JCYJ20130326110154156/JCYJ2015052509 2940984/ JCYJ20160422090807181)

Cartilage repair using induced pluripotent stem cell derived chondrocytes in osteoarthriti

Wu Zhang-song1, Zhu Hong-xia2, Luo Zhi-qiang1, Wu Xiao-min1, Lin Guang-yao1, Zhang Ming-yu1, Gu Hong-sheng2,Zhu Yan-xia1   

  1. 1Shenzhen Key Laboratory for Anti-ageing and Regenerative Medicine, Health Science Center, Shenzhen University, Shenzhen 518060, Guangdong Province, China; 2Department of Spinal Surgery, Xiaogan Maternity & Child Healthcare Hospital, Xiaogan 432100, Guangdong Province, China
  • Revised:2017-06-07 Online:2017-11-28 Published:2017-12-01
  • Contact: Zhu Yan-xia, M.D., Associate professor, Shenzhen Key Laboratory for Anti-ageing and Regenerative Medicine, Health Science Center, Shenzhen University, Shenzhen 518060, Guangdong Province, China
  • About author:Wu Zhang-song, Shenzhen Key Laboratory for Anti-ageing and Regenerative Medicine, Health Science Center, Shenzhen University, Shenzhen 518060, Guangdong Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81301597; Shenzhen Emerging Strategic Industrial Development Special Funds, No. JCYJ20130326110154156, JCYJ20150525092940984, and CYJ20160422090807181

摘要:

文章快速阅读:

文题释义:
诱导多能干细胞:
是由成纤维细胞经仙台病毒携带的4个反转录基因重编程而成的具有与胚胎干细胞类似生物学特性的多能干细胞。诱导多能干细胞具有很强的向软骨细胞分化的潜能,有望为临床骨关节炎发病机制的研究以及细胞治疗提供重要的细胞来源。
骨关节炎:是一种以关节软骨退行性变和继发性骨质增生为特性的慢性关节疾病,其发生主要与关节软骨细胞功能下降,软骨细胞外基质合成降解失衡有关,所以提供有功能的软骨细胞对于治疗骨关节炎是非常重要的。

 

摘要
背景:
基于干细胞的细胞治疗对于骨关节炎修复有重要的应用前景,其中诱导多能干细胞(induced pluripotent stem cell,iPS细胞)具有较强的增殖和分化潜能,且避免了胚胎干细胞的伦理学问题,目前被认为是细胞治疗中最有前景的种子细胞之一。
目的:拟探索一种iPS细胞向软骨细胞分化的有效方法,并研究iPS细胞来源软骨细胞对骨关节炎的修复作用。
方法:运用三步法将人iPS细胞诱导分化成软骨样细胞,并检测软骨细胞特异性基因和蛋白的表达。然后将分化后的iPS细胞移植到谷氨酸钠碘乙酸诱导的骨关节炎大鼠模型中。移植实验分为4组:对照组注射生理盐水,造模组注射谷氨酸钠碘乙酸,iPS细胞移植组注射谷氨酸钠碘乙酸后移植iPS细胞,分化后iPS细胞移植组注射谷氨酸钠碘乙酸后移植软骨分化后的iPS细胞。移植15周后micro-CT行断层扫描,并对移植后的关节进行组织学分析。
结果与结论:①与未分化的iPS细胞相比,经过6 d的拟胚体形成和2周的细胞分化,软骨细胞特异性基因和蛋白(Col2A1,GAG 和Sox9)的表达量都明显上升;②细胞移植15周后,未观察到免疫反应的发生,micro-CT显示移植细胞后软骨下骨的完整性明显得到改善,组织学分析也表明移植细胞后能产生关节软骨基质。iPS细胞来源软骨细胞的修复效果明显好于iPS细胞;③结果表明,iPS细胞来源软骨细胞移植可促进骨关节炎大鼠软骨再生,改善关节功能。

 

关键词: 干细胞, 移植, 诱导多能干细胞, 软骨细胞, 骨关节炎, 分化, 移植, 谷氨酸钠碘乙酸, 胶原, 蛋白多糖, Sox9, 国家自然科学基金

Abstract:

BACKGROUND: Stem cell-based therapy has been proposed for the treatment of osteoarthritis (OA) and induced pluripotent stem cells (iPSCs) are becoming a promising stem cell source as they have strong proliferation and differentiation potentials and no ethics problem. 
OBJECTIVE: To explore an effective method of iPSCs differentiating into chondrocytes and to study the therapeutic effect of iPSCs derived chondrocytes on osteoarthritis.
METHODS: In this study, three steps were developed to induce human iPSCs to differentiate into chondrocytes which were then transplanted into rat OA models induced by monosodium iodoacetate (MIA). There were four groups in the experiment: control group with normal saline injection, model group with MIA injection, iPSCs group with iPSCs transplantation following MIA injection, and differentiated iPSCs group with transplantation of iPSCs derived chondrocytes following MIA injection. At 15 weeks after transplantation, micro-CT was used and histological analysis of the knee joint was performed.
RESULTS AND CONCLUSION: Compared with the iPS group, the expression of chondrocyte specific genes and proteins (Col2A1, GAG and Sox9) were significantly increased in the differentiated iPSCs group after 6 days of embryoid formation and after 2 weeks of cell differentiation. At 15 weeks after cell transplantation, no immune responses were observed, micro-CT showed an improvement in subchondral bone integrity, and histological examinations demonstrated the production of articular cartilage matrix. iPSCs derived chondrocytes showed better effects on articular cartilage repair than the iPSCs. To conclude, iPSCs derived chondrocytes can be effective via transplantation approach for cartilage tissue regeneration in OA rats.

 

Key words: Osteoarthritis, Induced Pluripotent Stem Cells, Cell Differentiation, Chondrocytes, Tissue Engineering

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