Induced migration of endothelial cells into 3D scaffolds by chemoattractants secreted by pro-inflammatory macrophages in situ.
Induced migration of endothelial cells into 3D scaffolds by chemoattractants secreted by pro-inflammatory macrophages in situ.
复制标题
通过促炎巨噬细胞原位分泌的趋化剂诱导内皮细胞迁移到 3D 支架中
DOI：
10.1093/rb/rbx005
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发表时间：
2017-06
影响因子：
6.7
通讯作者：
Gao C
中科院分区：
文献类型：
--
作者：
Li X;Dai Y;Shen T;Gao C
摘要细胞在支架材料中的迁移是组织再生的关键，它能更好地模拟细胞在体内的行为。在这项研究中，已经提出了一种新的模型，控制三维细胞迁移的多孔胶原-壳聚糖支架与各种孔结构下的炎症细胞的刺激下，模拟血管生成过程。将Transwell模具置于24孔培养板的孔中，在支架顶部培养的内皮细胞（EC）被在孔培养板中孵育的促炎巨噬细胞分泌的化学引诱物如血管内皮生长因子（VEGF）和肿瘤坏死因子-α（TNF-α）促进迁移到支架中。50 ng/ml干扰素-γ（IFN-γ）和不同浓度的脂多糖（LPS，150-300 ng/ml）介导巨噬细胞表型。细胞迁移深度与LPS浓度呈正相关，与TNF-α浓度呈正相关。在-10 ° C（孔径为187 μm）下制备的支架中，内皮细胞比在-20 ° C（孔径为108 μm）下制备的支架更容易迁移到更深的区域。该方法提供了一个有用的策略来研究三维细胞迁移，并从长远来看，有助于揭示伤口愈合过程中的血管化过程。
Abstract Cell migration in scaffolds plays a crucial role in tissue regeneration, which can better mimic cell behaviors in vivo. In this study, a novel model has been proposed on controlling 3D cell migration in porous collagen-chitosan scaffolds with various pore structures under the stimulation of inflammatory cells to mimic the angiogenesis process. Endothelial cells (ECs) cultured atop the scaffolds in the Transwell molds which were placed into a well of a 24-well culture plate were promoted to migrate into the scaffolds by chemoattractants such as vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF-α) secreted by the pro-inflammatory macrophages incubated in the well culture plate. The phenotype of macrophages was mediated by 50 ng/ml interferon-gamma (IFN-γ) and different concentrations of lipopolysaccharide (LPS, 150–300 ng/ml). The cell migration depth had a positive correlation with LPS concentration, and thereby the TNF-α concentration. The ECs migrated easier to a deeper zone of the scaffolds prepared at − 10ºC (187 μm in pore diameter) than that at − 20ºC (108 μm in pore diameter) as well. The method provides a useful strategy to study the 3D cell migration, and is helpful to reveal the vascularization process during wound healing in the long run.