中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (5): 867-870.doi: 10.3969/j.issn.1673-8225.2012.05.026

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

传统及改良纱布包裹冻存法对人血管内皮细胞保护的比较*★

李  柳1,侯光辉2,吴  静3   

  1. 1暨南大学第一临床医院眼科,广东省广州市 510632;2暨南大学第三附属医院眼科,广东省广州市  519000;3暨南大学医学院眼科教研室,广东省广州市  510632
  • 收稿日期:2011-11-05 修回日期:2011-12-17 出版日期:2012-01-29
  • 通讯作者: 侯光辉,博士,硕士生导师,广东省广州市暨南大学第三附属医院眼科,广东省广州市 519000 houguanghui502@qq.com
  • 作者简介:李柳★,女,1988年生,江西省南昌市人,暨南大学在读硕士,主要从事角膜及眼表疾病方面的研究。liliu-0711@163.com
  • 基金资助:

    珠海市科技计划资助项目(PB20051014)。

Protective effects of traditional versus reforming cryopreserving methods on human vascular endothelial cells 

Li Liu1, Hou Guang-hui2, Wu Jing3   

  1. 1Department of Ophthalmology, the First Affiliated  Clinical Hospital of Jinan University, Guangzhou  510632, Guangdong Province, China; 2Department of Ophthalmology, the Third Affiliated Hospital of Jinan University, Guangzhou  519000, Guangdong Province, China; 3Department of Ophthalmology, Medical College of Jinan University, Guangzhou  510632, Guangdong Province, China
     
  • Received:2011-11-05 Revised:2011-12-17 Online:2012-01-29
  • Contact: Hou Guang-hui, Doctor, Master’s supervisor, Department of Ophthalmology, the Third Affiliated Hospital of Jinan University, Guangzhou 519000, Guangdong Province, China houguanghui502@qq.com
  • About author:Li Liu★, Studying for master’s degree, Department of Ophthalmology, the First Affiliated Clinical Hospital of Jinan University, Guangzhou 510632, Guangdong Province, China liliu-0711@163.com
  • Supported by:

    Zhuhai Municipal Science and Technology Program, No.PB20051014* 
       

摘要:

背景:传统的细胞冻存多采用4,-20 ℃放置的阶梯式降温方法,程序复杂且浪费时间。
目的:比较传统法与改良冻存法对血管内皮细胞的保护效果。
方法:将常规培养的人血管内皮细胞用胰酶消化后,用配置好的冻存液(10%二甲基亚砜、体积分数60%胎牛血清、30%DMEM无血清培养基)调整浓度,将细胞悬液吸入冻存管,分组干预:实验组以纱布包裹,并直接投入-80 ℃冰箱;对照组按常规冻存法,4 ℃冰箱和-20 ℃冰箱分别预冷30 min和1 h后放入-80 ℃冰箱。1个月后快速复苏冻存细胞。
结果与结论:复苏后实验组细胞存活率及生长曲线与对照组比较差异无显著性意义;复苏后实验组细胞贴壁率、形态学变化及增殖能力明显优于对照组。说明改良的纱布包裹法优于传统的细胞冻存法,且更加方便快捷。

关键词: 冻存, 纱布, 血管内皮细胞, MTT, 保护效果

Abstract:

BACKGROUND: As for traditional cryopreserving method, cells are cooling by steps at 4 ℃ and -20 ℃, which is complicated and a waste of time.
OBJECTIVE: To compare the protective effects of traditional and reforming cryopreserving methods on human vascular endothelial cells.
METHODS: The vascular endothelial cells were incubated with a freezing medium consisting of 10% dimethyl sulfoxide, 60% fetal bovine serum and 30% DMEM serum-free medium after digested with trypsin, and then the cell suspension were put into preserving tubes. Each group was intervened. In the experimental group, the tubes were packaged with gauze and then thawed into -80 ℃ refrigerator straightly. In the control group, the tubes were pre-cooling at 4 ℃ and -20 ℃ for 30 minutes and 1 hour, then thaw into -80 ℃ refrigerator. One month later, the endothelial cells were resuscitated.
RESULTS AND CONCLUSION: There were no significant difference in the survival rates and growth curves between experimental group and control group. The experimental group was significantly better than control group in adherence rates, morphological changes and proliferation. The reforming cryopreserving method is better than traditional cryopreserving method which is more convenient and easier to operate.

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