Identification of Skeletal Muscle Satellite Cells by Immunofluorescence with Pax7 and Laminin Antibodies

被引:16
|
作者
Feng, Xuesong [1 ]
Naz, Faiza [2 ]
Juan, Aster H. [1 ]
Dell'Orso, Stefania [2 ]
Sartorelli, Vittorio [1 ]
机构
[1] NIH, Lab Muscle Stem Cells & Gene Regulat, Bldg 10, Bethesda, MD 20892 USA
[2] Natl Inst Arthrit Musculoskeletal & Skin Dis NIAM, Off Sci & Technol, NIH, Bethesda, MD USA
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2018年 / 134期
基金
美国国家卫生研究院;
关键词
Developmental Biology; Issue; 134; Skeletal muscle; satellite cells; Pax7; immunofluorescence; stem cell; basal lamina; STEM-CELLS;
D O I
10.3791/57212
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Immunofluorescence is an effective method that helps to identify different cell types on tissue sections. In order to study the desired cell population, antibodies for specific cell markers are applied on tissue sections. In adult skeletal muscle, satellite cells (SCs) are stem cells that contribute to muscle repair and regeneration. Therefore, it is important to visualize and trace the satellite cell population under different physiological conditions. In resting skeletal muscle, SCs reside between the basal lamina and myofiber plasma membrane. A commonly used marker for identifying SCs on the myofibers or in cell culture is the paired box protein Pax7. In this article, an optimized Pax7 immunofluorescence protocol on skeletal muscle sections is presented that minimizes non-specific staining and background. Another antibody that recognizes a protein (laminin) of the basal lamina was also added to help identify SCs. Similar protocols can also be used to perform double or triple labeling with Pax7 and antibodies for additional proteins of interest.
引用
收藏
页数:6
相关论文
共 50 条
  • [31] Pax3 and Pax7 Exhibit Distinct and Overlapping Functions in Marking Muscle Satellite Cells and Muscle Repair in a Marine Teleost, Sebastes schlegelii
    Wang, Mengya
    Song, Weihao
    Jin, Chaofan
    Huang, Kejia
    Yu, Qianwen
    Qi, Jie
    Zhang, Quanqi
    He, Yan
    INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2021, 22 (07)
  • [32] PAX7 is a Sensitive Marker of Skeletal Muscle Differentiation in Rhabdomyosarcoma of the Female Gynecologic Tract
    Weiel, Julianna J.
    Kokh, Dina
    Charville, Gregory
    Longacre, Teri
    LABORATORY INVESTIGATION, 2020, 100 (SUPPL 1) : 1156 - 1156
  • [33] PAX7 target genes are globally repressed in facioscapulohumeral muscular dystrophy skeletal muscle
    Christopher R. S. Banerji
    Maryna Panamarova
    Husam Hebaishi
    Robert B. White
    Frédéric Relaix
    Simone Severini
    Peter S. Zammit
    Nature Communications, 8
  • [34] Pax7 directs postnatal renewal and propagation of myogenic satellite cells but not their specification
    Oustanina, S
    Hause, G
    Braun, T
    EMBO JOURNAL, 2004, 23 (16): : 3430 - 3439
  • [35] Detection of satellite cells during skeletal muscle wound healing in rats: time-dependent expressions of Pax7 and MyoD in relation to wound age
    Tian, Zhi-Ling
    Jiang, Shu-Kun
    Zhang, Miao
    Wang, Meng
    Li, Jiao-Yong
    Zhao, Rui
    Wang, Lin-Lin
    Li, Shan-Shan
    Liu, Min
    Zhang, Meng-Zhou
    Guan, Da-Wei
    INTERNATIONAL JOURNAL OF LEGAL MEDICINE, 2016, 130 (01) : 163 - 172
  • [36] Detection of satellite cells during skeletal muscle wound healing in rats: time-dependent expressions of Pax7 and MyoD in relation to wound age
    Zhi-Ling Tian
    Shu-Kun Jiang
    Miao Zhang
    Meng Wang
    Jiao-Yong Li
    Rui Zhao
    Lin-Lin Wang
    Shan-Shan Li
    Min Liu
    Meng-Zhou Zhang
    Da-Wei Guan
    International Journal of Legal Medicine, 2016, 130 : 163 - 172
  • [37] PAX7 is a Sensitive Marker of Skeletal Muscle Differentiation in Rhabdomyosarcoma of the Female Gynecologic Tract
    Weiel, Julianna J.
    Kokh, Dina
    Charville, Gregory
    Longacre, Teri
    MODERN PATHOLOGY, 2020, 33 (SUPPL 2) : 1156 - 1156
  • [38] microRNA-1 and microRNA-206 regulate skeletal muscle satellite cell proliferation and differentiation by repressing Pax7
    Chen, Jian-Fu
    Tao, Yazhong
    Li, Juan
    Deng, Zhongliang
    Yan, Zhen
    Xiao, Xiao
    Wang, Da-Zhi
    JOURNAL OF CELL BIOLOGY, 2010, 190 (05): : 867 - 879
  • [39] Sirtuin-1 Deacetylates Pax7 And Promotes Satellite Cell Differentiation In Injured Muscle
    Alway, Stephen E.
    Pitzer, Christopher R.
    Paez, Hector G.
    Sperringer, Justin E.
    Ferrandi, Peter J.
    Myers, Matthew J.
    Mohamed, Junaith S.
    MEDICINE AND SCIENCE IN SPORTS AND EXERCISE, 2021, 53 (08): : 117 - 117
  • [40] Insect peptide CopA3 promotes proliferation and PAX7 and MYOD expression in porcine muscle satellite cells
    Lee, Jeongeun
    Park, Jinryoung
    Choe, Hosung
    Shim, Kwanseob
    JOURNAL OF ANIMAL SCIENCE AND TECHNOLOGY, 2022, 64 (06) : 1132 - 1143