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  

Identification and analysis of chimeric rates in Down syndrome chimeric mice by quantitative real time PCR

作　　者：孟晓伟 杨冠恒[1,2] 王云杰 马晴雯[1,2] MENG Xiao-wei;YANG Guan-heng;WANG Yun-jie;MA Qing-wen(Shanghai Institute of Medical Genetics,Shanghai Children＇s Hospital,Shanghai Jiao Tong University,Shanghai 200040,China;Key Laboratory of Embryo MolecularBiology,Ministry of Health;Shanghai Key Laboratory of Embryo and Reproduction Engineering,Shanghai 200040,China)

机构地区：[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　　处：《上海交通大学学报（医学版）》2018年第11期1277-1282,共6页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家重点研发计划项目(2016YFC1000503)~~

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。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 calculated by detecting the relative quantity of human chromosome 21 （Hsa21） and mouse chromosome 15 （Mmu15） in samples. The relative quantity of Hsa21 and Mmu15 was evaluated with specifc primers for genes SIM2 on Hsa21 and Derl1 on Mmu15, respectively, by 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 detected by qPCR technology by using primers for genes on specifc 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.

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 

分 类 号：Q331[生物学—遗传学]