成纤维细胞是来源于胚胎中胚层的间充质细胞。由于成纤维细胞是最容易在体外培养的细胞类型之一,它们被广泛应用于各种细胞和分子研究。其耐久性使其适用于多种操作方法,包括基因转染研究和显微注射。组织中的成纤维细胞暴露于动态机械环境中,这影响着健康及愈合中的软组织的结构完整性。成纤维细胞分泌一种非刚性细胞外基质,富含 I 型和/或 III 型胶原。有证据表明,体内不同部位的成纤维细胞在内在特性上存在差异,并对周围细胞表现出不同的调控功能。例如,脾成纤维细胞已被证明可通过表达活性 IL-15 调节血液前体细胞向 NK 细胞的发育。
HSF 来自 ScienCell Research Laboratories,从人脾组织中分离而得。HSF 在传代一次后进行冷冻保存并以冷冻形式提供。每管含 >5 x 105 细胞,体积为 1 ml。HSF 以其梭形形态和对纤连蛋白特异抗体的免疫荧光特征进行表征。HSF 对 HIV-1、HBV、HCV、支原体、细菌、酵母和真菌均为阴性。HSF 在 ScienCell Research Laboratories 提供的条件下保证可进一步扩增 15 个世代倍增。
推荐培养基
建议使用成纤维细胞培养基(FM, Cat. #2301)进行体外培养 HSF。
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The spleen plays a critical role in the immune response against malaria parasite infection, where splenic fibroblasts (SFs) are abundantly present and contribute to immun... More
The spleen plays a critical role in the immune response against malaria parasite infection, where splenic fibroblasts (SFs) are abundantly present and contribute to immune function by secreting type I collagen (collagen I). The protein family is characterized by Plasmodium vivax tryptophan-rich antigens (PvTRAgs), comprising 40 members. PvTRAg23 has been reported to bind to human SFs (HSFs) and affect collagen I levels. Given the role of type I collagen in splenic immune function, it is important to investigate the functions of the other members within the PvTRAg protein family.
Methods
Protein structural prediction was conducted utilizing bioinformatics analysis tools and software. A total of 23 PvTRAgs were successfully expressed and purified using an Escherichia coli prokaryotic expression system, and the purified proteins were used for co-culture with HSFs. The collagen I levels and collagen-related signaling pathway protein levels were detected by immunoblotting, and the relative expression levels of inflammatory factors were determined by quantitative real-time PCR.
Results
In silico analysis showed that P. vivax has 40 genes encoding the TRAg family. The C-terminal region of all PvTRAgs is characterized by the presence of a domain rich in tryptophan residues. A total of 23 recombinant PvTRAgs were successfully expressed and purified. Only five PvTRAgs (PvTRAg5, PvTRAg16, PvTRAg23, PvTRAg30, and PvTRAg32) mediated the activation of the NF-κBp65 signaling pathway, which resulted in the production of inflammatory molecules and ultimately a significant reduction in collagen I levels in HSFs.
Conclusions
Our research contributes to the expansion of knowledge regarding the functional role of PvTRAgs, while it also enhances our understanding of the immune evasion mechanisms utilized by parasites. Less
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