上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2018, Vol. 38 ›› Issue (11): 1277-.doi: 10.3969/j.issn.1674-8115.2018.11.001

• 论著·基础研究 • 上一篇    下一篇

基于实时荧光定量PCR技术的唐氏综合征嵌合体小鼠嵌合率检测和分析

孟晓伟 1，杨冠恒 1, 2，王云杰 1, 2，马晴雯 1, 2   

  1. 1. 上海交通大学附属儿童医院，上海市儿童医院，上海交通大学医学遗传研究所，上海 200040；2. 卫生部医学胚胎分子生物学重点实验室暨上海市胚胎与生殖工程重点实验室，上海 200040
  • 出版日期:2018-11-28 发布日期:2018-12-15
  • 通讯作者: 马晴雯，电子信箱：maqingwen@hotmail.com。
  • 作者简介:孟晓伟（1992—），男，硕士生；电子信箱： xwmeng_@sjtu.edu.cn。
  • 基金资助:
    国家重点研发计划项目（2016YFC1000503）

Identification and analysis of chimeric rates in Down syndrome chimeric micequantitative real time PCR

MENG Xiao-wei1, YANG Guan-heng1, 2, WANG Yun-jie1, 2, MA Qing-wen1, 2   

  1. 1. Shanghai Institute of Medical Genetics, Shanghai Children’s Hospital, Shanghai Jiao Tong University, Shanghai 200040, China; 2. Key Laboratory of Embryo Molecular Biology, Ministry of Health; Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai 200040, China
  • Online:2018-11-28 Published:2018-12-15
  • Supported by:
    National Key Research and Development Program of China, 2016YFC1000503

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摘要： 目的 ·建立一种定量检测唐氏综合征嵌合体小鼠各脏器嵌合情况的方法，并初步探究其不同器官中的嵌合规律。方法 ·分别针对唐氏综合征模型 Tc1小鼠细胞（三体细胞）内人源 21号染色体（记为 Hsa21）上 SIM2基因和小鼠 15号染色体（记为 Mmu15）上 Derl1基因设计引物，采用实时荧光定量 PCR（quantitative real time PCR，qPCR）技术对 SIM2和 Derl1进行检测来反映 Hsa21和 Mmu15的比例，并以此计算嵌合体小鼠各器官中 Tc1小鼠细胞的嵌合占比。结果 ·所鉴定的小鼠中有 3只为嵌合体小鼠。该 3只小鼠心脏组织 Tc1小鼠细胞嵌合率分别为 8.98%、21.71%和 57.70%，小脑组织分别为 5.62%、20.17%和 40.43%，大脑组织分别为 8.48%、15.35%和 20.45%，肝脏组织分别为 2.66%、6.50%和 16.84%，脾脏组织分别为 1.73%、3.80%和 11.80%。结论 ·基于 qPCR技术对不同染色体上的基因进行定量分析的方法可用于唐氏综合征嵌合体小鼠中三体细胞嵌合率的定量检测。 Tc1小鼠细胞在嵌合体小鼠的心脏、小脑、大脑、肝脏、脾脏均可发生嵌合，心脏组织的嵌合率偏向最高。

关键词: 唐氏综合征, 嵌合体, 小鼠模型, 实时荧光定量 PCR, 嵌合率

Abstract:

Objective · To establish a method for identifying the chimeric rates in Down syndrome chimeric mice, and study the chimeric rules in different organs. Methods · The proportion of trisomic cells (Tc1 cells) in different organs was calculateddetecting the relative quantity of human chromosome 21 (Hsa21) and mochromosome 15 (Mmu15) in samples. The relative quantity of Hsa21 and Mmu15 was evaluated with specific primers for genes SIM2 on Hsa21 and Derl1 on Mmu15, respectively,quantitative real time PCR (qPCR) technology. Results · Three mice were chimeras and the chimeric levels of Tc1 cells were different in various tissues. The chimeric rates in hearts of these 3 mice were 8.98%, 21.71% and 57.70%; in cerebellums, the chimeric rates were 5.62%, 20.17% and 40.43%; in brains, the rates were 8.48%, 15.35% and 20.45%; in livers, the rates were 2.66%, 6.50% and 16.84%; and in spleens, the rates were 1.73%, 3.80% and 11.80%. Conclusion · The chimeric rate of trisomic cells in Down syndrome chimeric mice can be detectedqPCR technologyusing primers for genes on specific chromosomes. The chimerism occurs in the hearts, cerebellums, brains, livers and spleens of the chimeric mice and the chimeric rate of Tc1 cells tends to be the highest in the hearts.

Key words: Down syndrome, chimera, momodel, quantitative real time PCR, chimeric rate

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