spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

This Article
Right arrow Summary Freely available
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Fukada, S.-i.
Right arrow Articles by Yamamoto, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fukada, S.-i.
Right arrow Articles by Yamamoto, H.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

Muscle regeneration by reconstitution with bone marrow or fetal liver cells from green fluorescent protein-gene transgenic mice

So-ichiro Fukada1, Yuko Miyagoe-Suzuki2, Hiroshi Tsukihara1, Katsutoshi Yuasa2, Saito Higuchi1, Shiro Ono3, Kazutake Tsujikawa1, Shin'ichi Takeda2 and Hiroshi Yamamoto1,*

1 Department of Immunology, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan
2 Department of Molecular Therapy, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Kodaira, Tokyo 187-8502, Japan
3 Division of Oncogenesis, Biomedical Research Center, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan



View larger version (25K):

[in a new window]
  		Fig. 1. Experimental protocols for chimeras. For more details see the Materials and Methods section.

 


View larger version (55K):

[in a new window]
  		Fig. 2. Flow cytometric analyses of leukocyte chimerisms. (A) Splenocyte chimerisms of radiation-bone marrow chimera. (B) GFP+CD4+ and GFP+CD8+ T cells of the gated fraction of (A). (C) GFP+ cells in the peripheral blood of neonatal-bone-marrow chimera 4 weeks after reconstitution (4 weeks old). Splenic GFP+ cells (D) and splenic GFP+CD4+ and GFP+CD8+ T cells (E) of mouse C 9 weeks after reconstitution. (F) GFP+ bone marrow cells of mouse (C). (G) GFP+ cells in the peripheral blood of a neonatal-fetal liver cell chimera 4 weeks after reconstitution (4 weeks old). Thymic GFP+ cells (H), and CD4 and CD8 stainings of thymocytes (I) of mouse G at 10 weeks after reconstitution. Splenic GFP+ cells (J) and its GFP+CD4+ and GFP+CD8+ T cells (K) of mouse G. (L) GFP+ bone marrow cells of mouse G.

 


View larger version (103K):

[in a new window]
  		Fig. 3. Detection of GFP+ muscle fibres in the radiation bone marrow chimeras. H-E staining (A,C,E) and GFP fluorescence (B,D,F) of TA (A,B) and VL (C,D) muscles and diaphragm (E,F) of radiation-bone-marrow chimera. (G) GFP+ TA muscle fibre; (H) dystrophin staining (Texas-Red-conjugated anti-dystrophin antibody); (I) mixed color of (G) and (H); (J) GFP fluorescence; (K) dystrophin-staining; (L) the mixed color of (J) and (K) from the TA muscle of a normal C57BL/6 mouse as a control. Bars, 100 µm (A-F) and 50 µm (G-I).

 


View larger version (97K):

[in a new window]
  		Fig. 4. Detection of GFP+ muscle fibres and possible satellite cells in neonatal chimeras. H-E staining (A,C,E) and GFP fluorescence (B,D,F) of CTX-treated TA muscles of neonatal chimeras. (A,B) Neonatal bone marrow chimera; (C-F) neonatal fetal liver cell chimera; (G,H) a single GFP+ fibre with peripheral nucleus in a intact (CTX-untreated) neonatal bone marrow chimera; (I) detection of a GFP+ mononuclear cell beneath the laminin layer (rabbit anti-laminin plus rhodamine-conjugated goat anti-rabbit IgG) (neonatal bone marrow chimera); (J-O) desminstained explanted fibres (J,L,M,O) were examined for GFP fluorescence (J,K,M,N). A GFP+ cell attaching to a single fibre was stained with desmin (arrowheads of (L) and (O)). (P) Positive control of explanted muscle cells from GFP-Tg mouse (mixed color of desmin and GFP); (Q-T) are muscle sections of the secondary mdx/scid recipient. Single cell suspension from intact (CTX-untreated) muscles of neonatal bone marrow chimera was obtained and injected into CTX-treated TA muscle of the mdx/scid recipient. GFP+ fibres were observed (arrowheads of (R) and (T)). Bar, 50 µm (A-H and Q-T).

 


View larger version (13K):

[in a new window]
  		Fig. 5. Muscle-reconstituting efficiencies of bone marrow and fetal liver cells. Muscle-reconstituting efficiencies of neonatal bone marrow and fetal liver cells were compared as a factor of GFP+ muscle fibres/injected cell numbers. From the data shown in Table 1, both CTX-treated normal C57BL/6 and CTX-untreated mdx recipient mice were calculated.

 

Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?



© The Company of Biologists Ltd 2002