中国实验动物学报Issue(6):81-84,4.DOI:10.3969/j.issn.1005-4847.2014.06.014
Duchenne 型肌营养不良小鼠模型鉴定及干细胞移植后 dystrophin 的变化
Identification of the method of establishment of a DKO mouse model of Duchenne muscular dystrophy and regeneration of dystrophin expression in vivo after stem cell transplantation
摘要
Abstract
Objective To establish a method of identification of DKO mouse model of Duchenne muscular dystro-phy, and to assess the dystrophin regeneration after stem cell transplantation.Methods Heterozygous mice were mated and the resulting offspring were used to identify their genotype by SSP-PCR.The plasma creatine kinase level was measured by biochemical analyzer and histological changes in the DKO mice were analyzed using HE staining.Human umbilical cord mesenchymal stem cells were prepared and injected into the DKO mice hindlimb muscle, and dystrophin expression was de-tected by immunofluorescence staining at 2 months after injection.Results Mating of heterozygous mice generated three kinds of genotype offsprings, and 21.2%of the offsprings were identified as DKO genotype (285 bp) .DKO mice showed dystrophic symptoms, their plasma creatine kinase level was as high as 16988.52 ±617.48 IU/L, and significant histologi-cal changes including diverse myocyte sizes, numerous centrally nucleated cells and connective tissue proliferation or in-flammatory cells infiltration.Human dystrophin expression was detected in the DKO mouse hindlimb muscle at two months after injection of human umbilical cord mesenchymal stem cells.Conclusion DKO mouse genotype can be identified by SSP-PCR, and DKO mouse is an ideal animal model for studies of stem cell therapy for Duchenne muscular dystrophy.关键词
Duchenne型肌营养不良/动物模型/基因型鉴定/干细胞治疗/Dko小鼠Key words
Duchenne muscular dystrophy, Duchenne muscular dystrophy/Animal model/Genotype identifica-tion/Stem cell therapy/DkO mice分类
生物科学引用本文复制引用
庞荣清,张小飞,李自安,阮光萍,何洁,王强,王金祥,潘兴华,张成,张永云..Duchenne 型肌营养不良小鼠模型鉴定及干细胞移植后 dystrophin 的变化[J].中国实验动物学报,2014,(6):81-84,4.基金项目
云南省基金项目(2011HB050,2013DA004);成都军区基金项目(B12021,2012WJ003);“十二五”重大项目(2011ZXJ09201-031)。 ()