Stromal fibroblast-bone marrow-derived cell interactions: Implications for myofibroblast development in the cornea
- 作者：V. Singh ; V. Agrawal ; M.R. Santhiago ; S.E. Wilson ; wilsons4@ccf.org
- 关键词：corneal myofibroblasts ; corneal fibroblasts ; bone marrow-derived cells ; differentiation ; transforming growth factor beta ; paracrine ; juxtacrine ; green fluorescent protein ; alpha smooth muscle actin
- 刊名：Experimental Eye Research
- 出版年：2012
- 期刊代码：293_00144835
- 类别：med
- 出版时间：May, 2012
- 卷：98
- 期：Complete
- 页码：1-8
- 文件大小：1115 K
摘要
The purpose of this study was to test the hypothesis that mouse corneal stromal fibroblast and bone marrow-derived cell interactions augment corneal myofibroblast generation and, if so, to study whether such interactions are mediated by paracrine or juxtacrine mechanisms. Mouse bone marrow-derived cells and mouse corneal stromal fibroblasts were obtained from both mice with green fluorescent protein (GFP) expressed in all cells and normal GFP鈭?BL6 control mice. To study the interactions of the different cell types, GFP+ cells of one type were co-cultured with GFP鈭?cells of the other type in Primaria plates (to monitor juxtacrine signaling) or Transwell System plates (to monitor paracrine effects mediated by soluble mediators). Both cell types were cultured at a cell density of 1聽脳聽105聽cells聽per聽ml. The percentage of alpha smooth muscle actin+ myofibroblasts was significantly higher (ANOVA, p聽<聽0.001) when bone marrow-derived cells and mouse corneal stromal fibroblasts were co-cultured compared to when bone marrow-derived cells and mouse corneal stromal fibroblasts were cultured alone (control). The in聽vitro studies using GFP+ corneal fibroblasts or GFP+ bone marrow-derived cells demonstrated conclusively that both cells types could transform into myofibroblasts. However, the percentage of alpha smooth muscle actinassds+ myofibroblasts generated from either cell type precursor was higher when both cells were co-cultured together (juxtacrine) as compared to when bone marrow-derived cells and mouse corneal stromal fibroblasts were co-culture in different compartments of Transwell System (paracrine). Thus, more alpha smooth muscle actin+ GFP+ myofibroblasts were generated from GFP+ corneal stromal fibroblasts when GFP鈭?bone marrow-derived cells were present and more alpha smooth muscle actin+ GFP+ myofibroblasts were generated from GFP+ bone marrow-derived cells when GFP鈭?corneal stromal fibroblasts were present. Polyclonal anti-human latency associated peptide (LAP) (transforming growth factor-尾1) neutralizing antibody (a-LAP) and/or transforming growth factor-尾 type I receptor kinase inhibitor (LY-364947) inhibited the generation of alpha smooth muscle actin+ myofibroblasts from either precursor cell in Transwell System co-culture experiments. These data suggest that TGF尾 is a paracrine modulator that regulates the generation of myofibroblasts from either corneal fibroblasts or bone marrow-derived cell precursors.