牛骨骼肌卫星细胞的分离鉴定和诱导分化

Isolation, Identification and Induced Differentiation of Bovine Skeletal Muscle Satellite Cells

WANG Yi-min¹^，2, DAI Yang², LIU Xin-feng², LIU Zhong-wei², LI Ji-xia²,GUO Hong², DING Xiang-bin²

1.Institute of Bioengineering, Northwest A&F University, Yangling 712100, China；2. College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300384, China

Received:2013-12-17 Online:2014-07-20 Published:2014-08-21

Abstract

Abstract: In this study, bovine skeletal muscle satellite cells were isolated and identified, muscle tissues were digested by collagenase and trypsinase, then the satellite cells were purified and induced differentiate into myotubes. The muscle satellite cells and myotubes were observed, and detected by RT-PCR and immunofluorescent staining technology. The results showed that the muscle satellite cells were in healthy morphology, and more than 93 percent of the cells expressed the marker genes paired box protein 7(Pax7) and myogenic differentiation antigen(MyoD). After myogenesis induction, satellite cells began to convergence, and finally formed multinucleated myotubes expression of myogenin(MyoG) and myosin heavy chain(MHC). Our results indicated that high purity skeletal muscle satellite cells were successfully isolated and identification, which could provide cell source for muscle development and bovine muscle quality improvement study.

Key words:

bovine; skeletal muscle satellite cells; cell culture; cell differentiation; identification

WANG Yi-min, DAI Yang, LIU Xin-feng, LIU Zhong-wei, LI Ji-xia,GUO Hong, DING Xiang-bin. Isolation, Identification and Induced Differentiation of Bovine Skeletal Muscle Satellite Cells[J]. .

References

1 刘月光，史新娥，沈清武，等. 利用单根肌纤维法分离和培养猪骨骼肌卫星细胞及其成肌诱导分化［J］. 农业生物技术学报，2011，19(5):856～863.
2 庄新杰，严为巧，路易，等. 水牛骨骼肌卫星细胞的分离培养和生物学特性分析［J］. 黑龙江畜牧兽医，2009，11: 16～18.
3 张蔚菁，贾懿劼，田京. 肌卫星细胞增殖及分化影响因素研究进展［J］. 国际骨科学杂志，2013, 34(2): 104～106.
4 郑素月，樊宝良，张庆桥. 蛋鸡骨骼肌卫星细胞分离培养及鉴定［J］. 中国畜牧兽医，2011, 38(2): 80～83.
5 焦泽华，段彪，白春玲，等. 脱氧胆酸处理对大鼠肌肉卫星细胞的影响［J］. 中国畜牧兽医，2010, 37(12): 39～42.
6 Collins C A, Olsen I, Zammit P S, et al. Stem cell function, self-renewal, and behavioral heterogeneity of cells from the adult muscle satellite cell niche［J］. Cell, 2005, 122(2): 289～301.
7 Fukada S, Uezumi A, Ikemoto M, et al.Molecular signature of quiescent satellite cells in adult skeletal muscle［J］. Stem Cells, 2007, 25(10): 2448～2459.
8 Gizak A, Wrobel E, Moraczewski J, et al. Changes in subcellular localization of fructose 1,6-bisphosphatase during differentiation of isolated muscle satellite cells［J］. FEBS Lett, 2006, 580(17): 4042～4046.
9 Montarras D, Morgan J, Collins C, et al. Direct isolation of satellite cells for skeletal muscle regeneration［J］. Science, 2005, 309(5743): 2064～2067.
10 Pallafacchina G, Francois S, Regnault B, et al. An adult tissue specific stem cell in its niche: A gene profiling analysis of in vivo quiescent and activated muscle satellite cells［J］. Stem Cell Res, 2010, 4(2): 77～91.
11 Qu Z, Balkir L, van Deutekom J C, et al. Development of approaches to improve cell survival in myoblast transfer therapy［J］. The Journal of Cell Biology, 1998, 142(5): 1257～1267.
12 Sabourin L A， Rudnicki M A. The molecular regulation of myogenesis［J］. Clin Genet, 2000, 57(1): 16～25.
13 Sacco A, Doyonnas R, Kraft P, et al. Self-renewal and expansion of single transplanted muscle stem cells［J］. Nature, 2008, 456(7221): 502～506.
14 Seale P, Sabourin L A, Girgis-Gabardo A, et al. Pax7 is required for the specification of myogenic satellite cells［J］. Cell, 2000, 102(6): 777～786.
15 Starkey J D, Yamamoto M, Yamamoto S, et al. Skeletal muscle satellite cells are committed to myogenesis and do not spontaneously adopt nonmyogenic fates［J］. Journal Histochem Cytochem, 2011, 59(1): 33～46.
16 Zammit P S, Golding J P, Nagata Y, et al. Muscle satellite cells adopt divergent fates: A mechanism for self-renewal［J］. J Cell Biol, 2004, 166: 347～357.

Isolation, Identification and Induced Differentiation of Bovine Skeletal Muscle Satellite Cells

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 0

Recommended Articles

Metrics

Comments