Neutrophils Culture in Collagen Gel System.
Neutrophils Culture in Collagen Gel System.
复制标题
胶原凝胶系统中的中性粒细胞培养。
DOI：
10.3389/fimmu.2022.816037
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发表时间：
2022
影响因子：
7.3
通讯作者：
Ma Q
中科院分区：
文献类型：
--
作者：
Li R;Wang Z;Huang J;He S;Peng Y;Wan Y;Ma Q
中性粒细胞（Neu）能迅速迁移至受损组织，在宿主防御和组织稳态中起关键作用，包括肠上皮损伤和免疫应答。尽管它们在这些疾病中起着重要作用，但由于它们的寿命短且无法在体外冷冻保存或扩增，因此研究它们具有挑战性。此外，塑料板上的标准细胞培养（二维（2D）培养物）并不代表细胞驻留在组织中的实际微环境。在这项研究中，我们开发了一种新的三维（3D）培养系统的人和小鼠外周血Neu，这是由水凝胶。与2D培养系统相比，3D培养系统中的Neu显示出更好的细胞完整性和更少的细胞碎片。此外，与二维培养系统相比，三维培养系统更适合于观察经典刺激佛波酯（PMA）刺激的中性粒细胞胞外陷阱（NETs）以及脂多糖（LPS）/ATP、白细胞介素1 β（IL-1β）和肿瘤坏死因子α（TNFα）等损伤相关分子模式（DAMPs）。此外，三维培养体系中的NETs现象与体内相似。此外，对3D培养系统进行了评价，以共培养Neu和其他实质细胞，如结肠粘膜上皮细胞系。因此，三维培养系统较二维培养系统能更好地保持Neu的特性，并能缩短体内实验与体外实验之间的差距。
Neutrophils (Neu) migrate rapidly to damaged tissue and play critical roles in host defense and tissue homeostasis, including the intestinal epithelia injuries and immune responses. Although their important roles in these diseases, they are challenging to study due to their short life span and the inability to cryopreserve or expand them in vitro. Moreover, the standard cell culturing on plastic plates (two-dimensional (2D) cultures) does not represent the actual microenvironment where cells reside in tissues. In this study, we developed a new three-dimensional (3D) culture system for human and mouse peripheral blood Neu, which is made of hydrogel. The Neu showed much better cell integrity and less cell debris in the 3D culture system compared to that in 2D culture system. Moreover, the 3D culture system was more suitable for the observation of neutrophil extracellular traps (NETs) stimulated by the classical stimulation phorbol ester (PMA), and other damage associated molecular patterns (DAMPs) such as Lipopolysaccharide (LPS)/ATP, interleukin-1 β (IL-1β) and tumor necrosis factor α (TNFα) than the 2D culture system. Moreover, NETs phenomenon in 3D culture system is similar to that in vivo. In addition, the 3D culture system was evaluated to co-culturing Neu and other parenchymal cells, such as colon mucosal epithelial cell lines. In conclusion, the 3D culture system could maintain better properties of Neu than that in 2D culture system and it may reduce the gap between in vitro an in vivo experimentations.
影响因子：
7.3
作者：
Li Z;Peirasmaki D;Svärd S;Åbrink M
通讯作者：
Åbrink M