赵璐 付长振 赵志东等

摘要[目的]比较腺病毒和慢病毒侵染牛肌肉卫星细胞MOI值的差异，选择较优的侵染方式。[方法]从新生牛犊背最长肌中分离并纯化出牛肌肉卫星细胞，通过病毒侵染靶细胞的方法比较腺病毒和慢病毒对牛肌肉卫星细胞的侵染能力，确定最佳感染复数（Multiplicity of infection，MOI）。[结果]成功分离出牛肌肉卫星细胞，经过体外培养，可以用于细胞侵染研究；随着MOI值的增大，腺病毒和慢病毒侵染的细胞荧光强度均逐渐增强；当腺病毒侵染牛肌肉卫星细胞的MOI值为3 000，慢病毒侵染靶细胞的MOI值为300时，荧光强度均较强，细胞形态即将发生变化。相对而言，慢病毒较腺病毒更易侵染原代肌肉细胞，并且荧光强度较强，但侵染效率比较低。[结论]对于原代细胞，选择慢病毒侵染更为合适。该试验结果为应用腺病毒或慢病毒介导的功能基因在牛肌肉卫星细胞中的过表达或干扰沉默研究提供了参考资料。

关键词牛肌肉卫星细胞；腺病毒；慢病毒；MOI

中图分类号S858.23文献标识码A文章编号0517-6611（2016）04-147-04

Study on Adenovirus and Lentivirus Infection of Bovine Muscle Primary Cells

ZHAO Lu1， FU Changzhen2， ZHAO Zhidong2， LIU Yongfeng1* et al（1.College of Food Engineering and Nutritional Science， Shaanxi Normal University， Xian， Shaanxi 710062； 2. College of Animal Science and Technology， Northwest A&F University， Yangling， Shaanxi 712100）

Abstract[Objective] The aim was to compare the difference of MOI values between the adenovirus and the lentivirus infected bovine muscle satellite cells， and to select the best method of infection.[Method] Bovine muscle satellite cells were isolated and purified from newborn calf of longissimus muscle， infection ability of adenovirus and lentivirus to bovine muscle satellite cells were compared to determine the optimal MOI.[Result] Bovine muscle satellite cells were successfully isolated， which can be used for the study of cell infection by cultured in vitro. It was found that， with the increase of MOI values fluorescence intensity gradually increased for infected cells， the MOI infection of bovine muscle satellite cells was 3 000 for adenovirus with strong fluorescence. While， lentivirus infection of target cells MOI increased to 300 having strong fluorescence， and morphology will be changed. In comparison， lentivirus more easily than adenovirus infected primary cells， and the fluorescence intensity was stronger， but the infection efficiency was relatively low.[Conclusion] It is appropriate to select lentivirus infection of primary cells. The result provides reference for research of cattle genes function via using denovirus or lentivirus genes mediated bovine muscle satellite cells express or disturbanced in silence.

Key wordsBovine muscle satellite cells； Adenovirus； Lentivirus； MOI

骨骼肌衛星细胞是附着在骨骼肌肌纤维上对出生后骨骼肌生长和肌肉再生修复具有重要作用的一类干细胞[1]。激活后的肌卫星细胞即为肌肉前体细胞，经过多个回合的分裂，最后形成肌纤维[2]。肉牛不论是产肉量还是肉品质都与骨骼肌卫星细胞的数量、性质有密不可分的关系。出生以后的骨骼肌的生长主要依靠肌肉卫星细胞的补充，这些具有干细胞特性的细胞经过增殖和分化以后最终融合到已存在的肌纤维中[3]。……