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
  • [21] Regenerating human skeletal muscle forms an emerging niche in vivo to support PAX7 cells
    Hicks, Michael R.
    Saleh, Kholoud K.
    Clock, Ben
    Gibbs, Devin E.
    Yang, Mandee
    Younesi, Shahab
    Gane, Lily
    Gutierrez-Garcia, Victor
    Xi, Haibin
    Pyle, April D.
    NATURE CELL BIOLOGY, 2023, 25 (12) : 1758 - 1773
  • [22] Regenerating human skeletal muscle forms an emerging niche in vivo to support PAX7 cells
    Michael R. Hicks
    Kholoud K. Saleh
    Ben Clock
    Devin E. Gibbs
    Mandee Yang
    Shahab Younesi
    Lily Gane
    Victor Gutierrez-Garcia
    Haibin Xi
    April D. Pyle
    Nature Cell Biology, 2023, 25 : 1758 - 1773
  • [23] Modulation of Pax3 and Pax7 protein expression and localization in inactive skeletal muscle
    Hyatt, JPK
    Zhong, H
    Banker, TP
    Mikulich, MR
    Roy, RR
    FASEB JOURNAL, 2006, 20 (04): : A801 - A801
  • [24] Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line
    Yasuo Kitajima
    Yusuke Ono
    Skeletal Muscle, 8
  • [25] Pax7 distribution in human skeletal muscle biopsies and myogenic tissue cultures
    Reimann, J
    Brimah, K
    Schröder, R
    Wernig, A
    Beauchamp, JR
    Partridge, TA
    CELL AND TISSUE RESEARCH, 2004, 315 (02) : 233 - 242
  • [26] Pax7 distribution in human skeletal muscle biopsies and myogenic tissue cultures
    Jens Reimann
    Karima Brimah
    Rolf Schröder
    Anton Wernig
    Jonathan R. Beauchamp
    Terence A. Partridge
    Cell and Tissue Research, 2004, 315 : 233 - 242
  • [27] Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line
    Kitajima, Yasuo
    Ono, Yusuke
    SKELETAL MUSCLE, 2018, 8
  • [28] Pax7-expressing satellite cells are indispensable for adult skeletal muscle regeneration
    Sambasivan, Ramkumar
    Yao, Roseline
    Kissenpfennig, Adrien
    Van Wittenberghe, Laetitia
    Paldi, Andras
    Gayraud-Morel, Barbara
    Guenou, Hind
    Malissen, Bernard
    Tajbakhsh, Shahragim
    Galy, Anne
    DEVELOPMENT, 2011, 138 (17): : 3647 - 3656
  • [29] A key role for Pax7 transcripts in determination of muscle and nerve cells
    Ziman, MR
    Thomas, M
    Jacobsen, P
    Beazley, L
    EXPERIMENTAL CELL RESEARCH, 2001, 268 (02) : 220 - 229
  • [30] PAX7 target genes are globally repressed in facioscapulohumeral muscular dystrophy skeletal muscle
    Banerji, Christopher R. S.
    Panamarova, Maryna
    Hebaishi, Husam
    White, Robert B.
    Relaix, Frederic
    Severini, Simone
    Zammit, Peter S.
    NATURE COMMUNICATIONS, 2017, 8