中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (5): 809-812.doi: 10.3969/j.issn.1673-8225.2012.05.012

• 细胞与组织移植 cell and tissue transplantation • 上一篇    下一篇

新西兰雌-雄兔皮肤移植模型体内细胞毒性实验*★

梁伟潮,蒋泽生,汪  燕,钟利民,潘明新   

  1. 南方医科大学附属珠江医院肝胆二科,广东省广州市  515282
  • 收稿日期:2011-07-04 修回日期:2011-10-24 出版日期:2012-01-29
  • 通讯作者: 潘明新,教授,南方医科大学附属珠江医院肝胆二科,广东省广州市 515282 pmxwxy@sohu.com
  • 作者简介:梁伟潮★,男,1979年生,广东省佛山市人,汉族,2003年中山大学毕业,硕士,主治医师,主要从事移植免疫研究。 lwchng@qq.com
  • 基金资助:

    国家自然科学基金资助项目(30972825)“供体抗原特异性体内免疫状态体内定量检测的基础研究”。

Cytotoxicity assay of New Zealand female-male rabbit skin graft models in vivo

Liang Wei-chao, Jiang Ze-sheng, Wang Yan, Zhong Li-min, Pan Ming-xin   

  1. Second Department of Hepatobiliary Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou  515282, Guangdong Province, China
  • Received:2011-07-04 Revised:2011-10-24 Online:2012-01-29
  • Contact: Pan Ming-xin, Professor, Second Department of Hepatobiliary Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 515282, Guangdong Province, China pmxwxy@sohu.com
  • About author:Liang Wei-chao★, Master, Attending physician, Second Department of Hepatobiliary Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China lwchng@qq.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30972825*

摘要:

背景:建立一种有效的评估模式来监测移植后受者免疫状态是当前的难点。
目的:建立兔体内细胞毒性实验方法,验证体内细胞毒性实验能否检测出受体的免疫排斥状况。
方法:建立新西兰白兔雌-雄兔皮肤移植模型,另设未作移植的雌、雄兔为对照组。取雌兔及雄兔脾脏制备脾细胞单细胞悬液,分别用不同浓度活细胞染料羟基荧光素乙酰乙酸琥珀酰亚胺酯染色后制备1∶1混合细胞悬液。雄兔输注混合细胞后分别于1,2,4,8 h 抽取外周血,流式细胞仪检测外周血两种荧光细胞的比例变化,计算供体特异性细胞杀伤率。
结果与结论:雌-雄兔皮肤移植排斥模型在皮肤移植2周后可建立,混合细胞悬液输注后在模型组和对照组外周血中的比例变化不同,雄兔输注混合细胞后1,2,4,8 h的杀伤率模型组高于对照组,差异有显著性意义(P < 0.01)。提示体内细胞毒性实验可用于免疫状态监测,可以直观地反映皮肤移植模型的体内特异性免疫环境及移植物所受的免疫攻击强度。

关键词: 细胞毒性实验, 移植排斥, 免疫监测, 流式细胞术, 皮肤移植

Abstract:

BACKGROUND: The establishment of an effective assessment model for monitoring post-transplant recipient's immune status is the difficulty in current researches.
OBJECTIVE: To establish an in vivo method of cytotoxicity assay in rabbits; to verify whether the in vivo cytotoxicity assay can detect the immune status of the recipient rabbits.
METHODS: New Zealand female-male rabbit skin graft models were established; the untransplanted female and male rabbits served as control. The spleens of both female and male rabbits were removed to prepare single cell suspension; and then live cell dye hydroxyfluorescein diacetate succinimidyl ester of different concentrations were added to stain and prepare for the 1:1 mixed cell suspension. Peripheral blood samples of the male rabbits were collected at 1, 2, 3 and 8 hours after the infusion of the mixed cell suspension. Flow cytometry was used to detect the ratio of the two kinds of fluorescytes in the peripheral blood; the killing rate of specific cells in the donors was calculated.
RESULTS AND CONCLUSION: The female-male rabbit model of skin graft rejection was established in the 2nd week after skin transplantaion. The ratio changes in the peripheral blood of model group and control group were different after the infusion of mixed cell suspension; the killing rate in the male rabbits of the model group was significantly higher than that of the control group at 1, 2, 4 and 8 hours after infusion (P < 0.01). These findings indicate that cytotoxicity assay in vivo can be used to monitor immune state; it can directly reflect the specific immune environment in vivo of the skin graft models and the intensity of the immune attack that the graft suffered.

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