Direct isolation of myofibroblasts and fibroblasts from bleomycin-injured lungs reveals their functional similarities and differences

Fibrogenesis & Tissue Repair

Toshihide Iwashita⁴  Kingo Chida¹  Takafumi Suda¹  Kiyoshi Shibata³  Makiko Sakao⁴  Shiori Meguro⁴  Hideya Kawasaki⁴  Isao Kosugi⁴  Yosifumi Arai²  Taisuke Akamatsu⁴ 

[1] Department of Respiratory Medicine, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Japan;Department of Pathology, Seirei Hamamatsu General Hospital, 2-12-12 Sumiyoshi, Naka-ku, Hamamatsu, Japan;Research Equipment Center, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Japan;Department of Regenerative and Infectious Pathology, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Japan

关键词: Myofibroblasts;    Fibroblasts;    Collagen;    Chemokine;    α-smooth muscle actin;   

Others  :  803012
DOI  :  10.1186/1755-1536-6-15

 received in 2013-04-10, accepted in 2013-07-08,  发布年份 2013

【 摘 要 】

Background

Myofibroblasts play a crucial role in tissue repair. The functional similarities and differences between myofibroblasts and fibroblasts are not fully understood because they have not been separately isolated from a living body. The purpose of this study was to establish a method for the direct isolation of myofibroblasts and fibroblasts from injured lungs by using fluorescence-activated cell sorting and to compare their functions.

Results

We demonstrated that lineage-specific cell surface markers (lin), such as CD31, CD45, CD146, EpCAM (CD326), TER119, and Lyve-1 were not expressed in myofibroblasts or fibroblasts. Fibroblasts of bleomycin-injured lungs and saline-treated lungs were shown to be enriched in lin^neg Sca-1^high, and myofibroblasts of bleomycin-injured lungs were shown to be enriched in lin^neg Sca-1^low CD49e^high. Results from in-vitro proliferation assays indicated in-vitro proliferation of fibroblasts but not myofibroblasts of bleomycin-injured lungs and of fibroblasts of saline-treated lungs. However, fibroblasts and myofibroblasts might have a low proliferative capacity in vivo. Analysis of genes for collagen and collagen synthesis enzymes by qRT-PCR showed that the expression levels of about half of the genes were significantly higher in fibroblasts and myofibroblasts of bleomycin-injured lungs than in fibroblasts of saline-treated lungs. By contrast, the expression levels of 8 of 11 chemokine genes of myofibroblasts were significantly lower than those of fibroblasts.

Conclusions

This is the first study showing a direct isolation method of myofibroblasts and fibroblasts from injured lungs. We demonstrated functional similarities and differences between myofibroblasts and fibroblasts in terms of both their proliferative capacity and the expression levels of genes for collagen, collagen synthesis enzymes, and chemokines. Thus, this direct isolation method has great potential for obtaining useful information from myofibroblasts and fibroblasts.

【 授权许可】

   
2013 Akamatsu et al.; licensee BioMed Central Ltd.

【 图 表 】

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