中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (47): 8783-8786.doi: 10.3969/j.issn.1673-8225.2010.47.011

• 组织工程血管材料 tissue-engineered vascular materials • 上一篇    下一篇

人脐静脉脱细胞基质的制备

王海江1，范应中2，李泸平2，张  震2，张  大2   

  1. 1深圳市第三人民医院外科，广东省深圳市   518020；2郑州大学第一附属医院小儿外科，河南省郑州市   450052　
  • 出版日期:2010-11-19 发布日期:2010-11-19
  • 通讯作者: 范应中，博士，教授，郑州大学第一附属医院小儿外科，河南省郑州市 450052 fanyingzhong@126.com
  • 作者简介:王海江★，男，1973年生，河南省虞城县人，汉族，2009年郑州大学第一临床学院毕业，硕士，主治医师，主要从事组织工程方面的研究。 Whaj2008@163.com

Preparation of extracellular matrix of human umbilical veins

Wang Hai-jiang1, Fan Ying-zhong2, Li Lu-ping2, Zhang Zhen2, Zhang Da2   

  1. 1 Department of Surgery, The Third People’s Hospital of Shenzhen, Shenzhen   518020, Guangdong Province, China; 2 Department of Pediatric Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou   450052, Henan Province, China 　
  • Online:2010-11-19 Published:2010-11-19
  • Contact: Fan Ying-zhong, Doctor, Professor, Department of Pediatric Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China fanyingzhong@126.com
  • About author:Wang Hai-jiang★, Master, Attending physician, Department of Surgery, The Third People’s Hospital of Shenzhen, Shenzhen 518020, Guangdong Province, China Whaj2008@163.com

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摘要：

背景：细胞外基质的制备以去除组织中的细胞成分为目的，目前脱细胞方法尚无统一的标准，但是制备的细胞外基质均需证实有无细胞残留。
目的：观察应用酶消化、去污剂、渗透溶液方法和TritonX-100与氨水混合液方法制备脐静脉脱细胞基质的效果，以探讨制备人脐静脉脱细胞基质的理想方法。
方法：分别采用酶消化、去污剂、渗透溶液法和曲拉松与氨水混合液两种方法处理人脐静脉，制备人脐静脉脱细胞基质。观察脱细胞处理前后脐静脉基质的物理性状和苏木精-伊红染色结果，随机观察50个400倍镜像下残留细胞碎片数目。
结果与结论：脐静脉经脱细胞处理后呈瓷白色半透明胶冻样管状结构。酶消化、去污剂、渗透溶液方法组的人脐静脉细胞成分几乎全部去除，细胞外基质保持完好，纤维结构完整，曲拉松与氨水混合液方法组有不同数量的残余细胞成分，两组差异具有显著性意义(P < 0.05)。结果表明，酶消化、去污剂和渗透溶液法是制备人脐静脉脱细胞基质较为理想的方法。

关键词: 细胞外基质, 人脐静脉, 制备, 组织工程, 酶消化, 去污剂, 渗透溶液

Abstract:

BACKGROUND: Preparation of extracellular matrix (ECM) aims to remove the cellular components in tissues, there is no uniform standard for the current acellular methods, but the prepared ECM should be proved to be without residual cells. 
OBJECTIVE: To prepare human umbilical vein ECM by using hypotonic and hypertonic solutions, detergent and proteinase in a multistep process, mixed solution of Triton X-100 and ammonia water, and to explore the ideal method of preparing human umbilical vein ECM. 
METHODS: ECM of human umbilical veins was prepared by use of hypotonic and hypertonic solutions, detergent and proteinase in a multistep process and solution of Triton X-100 and NH3·H2O. The physical properties and hematoxylin-eosin staining of human umbilical vein before and after acellular matrix were observed, the amount of residual cell debris in 50 images at 400 magnification was measured.
RESULTS AND CONCLUSION: The acellular umbilical veins exhibited translucent porcelain-white jelly-like tubular structure. The cell components of the human umbilical veins resulting from hypotonic and hypertonic solutions, detergent and proteinase were almost removed, ECM maintained an integrity, and fibrous structures were preserved. ECM resulting from solution of Triton-X100 and NH3·H2O had different quantities of remaining cellular elements. There were significant differences between the two groups (P < 0.05). Hypotonic and hypertonic solutions, detergent and proteinase are ideal methods to prepare ECM from human umbilical veins.

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