中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (40): 7505-7508.doi: 10.3969/j.issn.1673-8225.2010.40.021

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

双细胞种植改善人工血管内皮细胞的保留率

裴  斐，何  蕊，李俊彦，张  莉，陈  旭   

  1. 西安交通大学第二附属医院心血管外科，陕西省西安市  710004
  • 出版日期:2010-10-01 发布日期:2010-10-01
  • 作者简介:裴斐★，男，1964年生，陕西省西安市人，汉族，1986年西安医科大学毕业，硕士，副教授，主要从事生物医学工程和心血管外科学方面的研究。 menglan6-312@126.com
  • 基金资助:

    国家自然科学基金资助项目(30470456)，zg-tPA/eNOS基因修饰/双细胞种植改善小口径人工血管通畅率的实验研究。　

Double layer cells implantation improves retention rate of vascular prosthesis endothelial cells

Pei Fei, He Rui, Li Jun-yan, Zhang Li, Chen Xu   

  1. Department of Cardiovascular Surgery, Second Affiliated Hospital, Xi’an Jiaotong University, Xi’an  710004, Shaanxi Province, China
  • Online:2010-10-01 Published:2010-10-01
  • About author:Pei Fei★, Master, Associate professor, Department of Cardiovascular Surgery, Second Affiliated Hospital, Xi’an Jiaotong University, Xi’an 710004, Shaanxi Province, China menglan6-312@126.com
  • Supported by:

    the  National Natural    Science Foundation of China, No. 30470456*

PDF

483

被引次数

6

摘要：

背景：前期研究表明，在人工血管内壁种植内皮细胞，使其早期快速内皮化，可以改善移植血管的通畅性，但种植的单层内皮细胞往往黏附力差，在血流的冲击下容易脱落。
目的：采用血管内皮细胞和平滑肌细胞双层种植方法，提高人工血管腔面内皮细胞的保存率。
方法：采用标准酶分离技术和细胞培养技术分离培养兔内皮细胞。采用组织贴块法分离培养兔平滑肌细胞。并分别以lacZ和GFP基因转染内皮细胞及平滑肌细胞。双层细胞种植组在聚四氟乙烯人工血管管腔面先后种植平滑肌细胞和内皮细胞，以单纯种植血管内皮细胞为对照。在体外将受试的人工血管接入作者设计的灌流模型下灌注2 h，计算比较灌注前后受试标本内皮细胞密度。体内实验，将人工血管植入兔子体内7 d，分别测定内皮细胞保留率。
结果与结论：灌注2 h 后，单纯内皮细胞种植组细胞保留率为0.39±0.04，而双层细胞种植组内皮细胞保留率为0.73±0.07，双层细胞种植组细胞保存率明显高于单纯内皮细胞种植组(P < 0.01)。人工血管植入兔体内7 d，单纯内皮细胞种植组内皮细胞保留率为0.63±0.10，双层细胞种植组内皮细胞保留率为0.95±0.06，两组间差异有显著性意义(P < 0.05)。结果证实在内皮细胞和人工血管壁之间种植平滑肌细胞在体外和体内均可提高内皮细胞的保存率。

关键词: 人工血管, 血管内皮细胞, 血管平滑肌细胞, 细胞种植, 保留率　

Abstract:

BACKGROUND: Previous studies have shown that vascular endothelial cells were seed in grafts to enhance endothelialization rapidly and early, which can improve graft fluency. But the adhesion of seeded endothelial cells is often poor in single-layer. They are easily washed off under the blood flow.
OBJECTIVE: To improve cell retention on the graft with dual cell seeding technique, adding a layer of smooth muscle cells (SMCs) between the graft and vascular endothelial cells.
METHODS: Rabbit endothelial cells were cultured by standard enzyme separation technique and cell culture technique. Rabbit smooth muscle cells were isolated and cultured using tissue explant method. The lacz and green fluorescent porotein genes were used to transfer endothelial cells and smooth muscle cells. Polytetrafluoroethylene grafts pretreated with fibronectin were seeded with smooth muscle cell followed by endothelial cells in two-layer cells group. Vascular endothelial cells implantation served as controls. Vascular prosthesis was connected to perfusion models for 2 hours. The density of endothelial cells was calculated prior to and following perfusion. In vivo experiment, cell retention rate was measured following vascular prosthesis was inserted into the rabbit for 7 days.
RESULTS AND CONCLUSION: 2 hours following perfusion, cell retention rate was (0.39±0.04) in endothelial cells group, and (0.73±0.07) in the two-lay cells group. The cell retention rate was greater in the two-layer cells group compared with endothelial cells group (P < 0.01). 7 days following vascular prosthesis implantation, the cell retention rate was (0.63±0.10) in the endothelial cells group, and (0.95±0.06) in the two-layer cells group (P < 0.05). Results indicated that using smooth muscle cell as a media layer between endothelial cells and graft surface can improve the retention of seeded endothelial cells in vitro and vivo.

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