多浪绵羊骨骼肌卫星细胞的分离培养、鉴定及成肌诱导分化

doi:10.16431/j.cnki.1671-7236.2016.11.022

Abstract

Abstract:

To mimic muscle development of Duolang sheep in vitro,we employed a two-step digestion method to separate satellite cells（SCs）and a differential adhesion method to purify the cells in Duolang sheep.Moreover,observation of microscopic images and immunofluorescence were used for identifying Duolang sheep SCs and its myogenic differentiation.Using immunofluorescence for Desmin,Pax7 and MyoD1 genes,we demonstrated that these marker genes all expressed in the SCs.The SCs formed significant myotubes when the serum was withdrawal from growth media,confirmed by the immunofluorescence for MHC and microscopic images.Taken together,we ssuccessfully isolated SCs and established the myogenic differentiation of SCs.

Key words: sheep; skeletal muscle satellite cell; isolation and culture; myogenic differentiation

CLC Number:

Q813

ZHAO Qian, ZHANG Ping, LI Xiang-chen, GUAN Wei-jun, PU Ya-bin, ZHAO Qian-jun, MA Yue-hui. Isolation,Culture,Identification and Myogenic Differentiation of Skeletal Muscle Satellite Cells in Duolang Sheep[J]. , 2016, 43(11): 2956-2962.

References

[1] Relaix F,Zammit P S.Satellite cells are essential for skeletal muscle regeneration:The cell on the edge returns centre stage[J].Development,2012,139(16):2845-2856.
[2] 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.
[3] Wu H,Ren Y,Li S,et al.In vitro culture and induced differentiation of sheep skeletal muscle satellite cells[J].Cell Biol Int,2012,36(6):579-587.
[4] Moresi V,Marroncelli N,Adamo S.New insights into the epigenetic control of satellite cells[J].World J Stem Cells,2015,7(6):945-955.
[5] 沈林園,张顺华,吴泽辉,等.骨骼肌卫星细胞对肉品质的影响及其分化调控[J].遗传,2013,35(9):1081-1086.
[6] Murphy M M,Lawson J A,Mathew S J,et al.Satellite cells,connective tissue fibroblasts and their interactions are crucial for muscle regeneration[J].Development,2011,138(17):3625-3637.
[7] 李方华,弓慧敏,冯士彬,等.骨骼肌卫星细胞的研究进展[J].中国畜牧兽医,2010,37(10):25-28.
[8] 陈岩,王琨,朱大海.鸡骨骼肌卫星细胞的分离培养,鉴定及生物学特性研究[J].遗传,2006,28(3):257-260.
[9] 李方华,侯玲玲,马月辉,等.北京油鸡骨骼肌卫星细胞的分离,培养,鉴定及成肌诱导分化的研究[J].中国农业科学,2010,43(22):4725-4731.
[10] 王轶敏,代阳,刘新峰,等.牛骨骼肌卫星细胞的分离鉴定和诱导分化[J].中国畜牧兽医,2014,41(7):142-147.
[11] 林凯,虞德兵,解晓东,等.TGF-β/Smad信号通路在Follistatin调节鸭骨骼肌卫星细胞增殖过程中的作用机制[J].中国农业科学,2015,48(12):2460-2468.
[12] 何波,郑嵘,熊远著,等.新生猪骨骼肌卫星细胞的培养鉴定及生物学特性[J].畜牧兽医学报,2006,37(6):555-559.
[13] 边艳超,牧仁,李向臣,等.鲁西黄牛胚胎骨骼肌卫星细胞分离培养及生物学特性[J].华中农业大学学报,2013,32(2):90-96.
[14] 王慧华,赵福平,张莉,等.中国地方绵羊品种的地域分布及肉用相关性状的多元分析[J].中国农业科学,2015,48(20):4170-4177.
[15] Dick S A,Chang N C,Dumont N A,et al.Caspase 3 cleavage of Pax7 inhibits self-renewal of satellite cells[J].Proc Natl Acad Sci USA,2015,112(38):E5246-E5252.
[16] Xiao L,Lee K K.BRE facilitates skeletal muscle regeneration by promoting satellite cell motility and differentiation[J].Biol Open,2016,5(2):100-111.
[17] Hindi S M,Kumar A.TRAF6 regulates satellite stem cell self-renewal and function during regenerative myogenesis[J].J Clin Invest,2015,126(1):152-168.
[18] Wu R,Li H,Zhai L,et al.microRNA-431 accelerates muscle regeneration and ameliorates muscular dystrophy by targeting Pax7 in mice[J].Nat Commun,2015,6:7713.
[19] 杨伟钧,王修启,王松波,等.骨骼肌卫星细胞自我更新的研究进展[J].中国畜牧兽医,2010,40(8):101-106.
[20] Brack A S,Rando T A.Tissue-specific stem cells:Lessons from the skeletal muscle satellite cell[J].Cell Stem Cell,2012,10(5):504-514.
[21] Tierney M T,Aydogdu T,Sala D,et al.STAT3 signaling controls satellite cell expansion and skeletal muscle repair[J].Nat Med,2014,20(10):1182-1186.
[22] Xu X,Wilschut K J,Kouklis G,et al.Human satellite cell transplantation and regeneration from diverse skeletal muscles[J].Stem Cell Reports,2015,5(3):419-434.

Isolation,Culture,Identification and Myogenic Differentiation of Skeletal Muscle Satellite Cells in Duolang Sheep

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