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腹膜间皮下基质细胞支持造血并分化为成骨和成脂细胞谱系。

Peritoneal Submesothelial Stromal Cells Support Hematopoiesis and Differentiate into Osteogenic and Adipogenic Cell Lineages.

作者信息

do Amaral R J F C, Benac P, Andrade L R, Farina M, Bernardazzi C, Arcanjo K D, Palumbo A, Cordeiro I R, Brito J M, El-Cheikh M C, Oliveira F L

机构信息

Laboratório de Proliferação e Diferenciação Celular, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.

出版信息

Cells Tissues Organs. 2014;200(2):118-31. doi: 10.1159/000377624. Epub 2015 May 9.

Abstract

The peritoneum is a thin membrane that covers most of the abdominal organs, composed of a monolayer of mesothelial cells and subjacent submesothelial loose connective tissue. Cells from the peritoneal wall are correlated with peritoneal fibrosis and epithelial-to-mesenchymal transition. However, the distinct involvement of mesothelial or submesothelial cells in such phenomena is still not clear. Here, we propose a new strategy to obtain stromal cells from anterior peritoneal wall explant cultures. These cells migrated from peritoneal tissues and proliferated in vitro for 4 weeks as adherent fibroblast-like cells. Optical and electronic microscopy analyses of the fragments revealed a significant submesothelial disorganization. The obtained cells were characterized as cytokeratin- vimentin+ laminin+ α-smooth muscle actin+, suggesting a connective tissue origin. Moreover, at the third passage, these stromal cells were CD90+CD73+CD29+Flk-1+CD45-, a phenotype normally attributed to cells of mesenchymal origin. These cells were able to support hematopoiesis, expressing genes involved in myelopoiesis (SCF, G-CSF, GM-CSF, IL-7 and CXCL-12), and differentiated into osteogenic and adipogenic cell lineages. The methodology demonstrated in this work can be considered an excellent experimental model to understand the physiology of the peritoneal wall in healthy and pathological processes. Moreover, this work shows for the first time that submesothelial stromal cells have properties similar to those of mesenchymal cells from other origins.

摘要

腹膜是一层覆盖大部分腹腔器官的薄膜,由单层间皮细胞和下方的间皮下疏松结缔组织组成。腹膜壁细胞与腹膜纤维化和上皮-间充质转化相关。然而,间皮细胞或间皮下细胞在这些现象中的具体作用仍不清楚。在此,我们提出了一种从腹膜前壁外植体培养物中获取基质细胞的新策略。这些细胞从腹膜组织迁移而来,并在体外作为贴壁的成纤维细胞样细胞增殖4周。对这些组织碎片的光学和电子显微镜分析显示间皮下有明显的结构紊乱。所获得的细胞被鉴定为细胞角蛋白阳性、波形蛋白阳性、层粘连蛋白阳性、α-平滑肌肌动蛋白阳性,提示其起源于结缔组织。此外,在第三代时,这些基质细胞为CD90阳性、CD73阳性、CD29阳性、Flk-1阳性、CD45阴性,这种表型通常归因于间充质来源的细胞。这些细胞能够支持造血,表达参与骨髓生成的基因(干细胞因子、粒细胞集落刺激因子、粒细胞-巨噬细胞集落刺激因子、白细胞介素-7和CXC趋化因子配体12),并分化为成骨和成脂细胞谱系。这项工作中展示的方法可被视为理解腹膜壁在健康和病理过程中的生理学的一个优秀实验模型。此外,这项工作首次表明间皮下基质细胞具有与其他来源的间充质细胞相似的特性。

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