Transmigration of macrophages through primary adult rat Sertoli cells.

IF 3.6 Q2 MEDICINE, RESEARCH & EXPERIMENTAL Tissue Barriers Pub Date : 2023-01-02 DOI:10.1080/21688370.2022.2064179
Hassan Kabbesh, Muhammad A Riaz, Alexandra D Jensen, Georgios Scheiner-Bobis, Lutz Konrad
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引用次数: 1

Abstract

The blood testis barrier (BTB) is often studied with isolated immature Sertoli cells (SCs), transepithelial resistance (TER) measurements and FITC dextran diffusion assays. Recently, it was found that even in the absence of SCs, only few immune cells enter the seminiferous tubules. Thus, in this study, we evaluated the testicular immunological barrier (TIB) in vitro by transmigration of macrophages through SCs with and without peritubular cells (PCs) and with or without matrigel (MG). Primary PCs were isolated from adult rat testis and kept in mono- or co-cultures with the conditionally reprogrammed primary adult Sertoli cell line (PASC1) from rat that has been recently generated by our group. Rat monocytes isolated from fresh blood were differentiated into M0 macrophages, and after polarization to M1 or M2 macrophages characterized by gene expression of CXCL11 and TNF-α for M1, or CCL17 and CCL22 for M2. Transmigration of LeukoTracker-labeled M0, M1, and M2 macrophages through mono- and co-cultures of PCs/SCs with and without MG demonstrated that SCs are the main constituent of the TIB in vitro with only a negligible contribution of PCs or MG. Moreover, M2 macrophages showed less migration activity compared to M0 or M1. Treatment of SCs with testosterone (T) showed positive effects on the barrier in contrast to negative effects by interleukin-6 (IL-6) or tumor necrosis factor-α (TNF-α). The new transmigration model is suitable to evaluate transmigration of macrophages through a barrier consisting of testicular cells and can be applied to study the integrity of testicular barriers with respect to immunological aspects.

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巨噬细胞在成年大鼠原代支持细胞中的迁移。
血睾丸屏障(BTB)通常通过分离的未成熟支持细胞(SCs)、经上皮耐药(TER)测量和FITC葡聚糖扩散试验来研究。最近发现,即使在没有SCs的情况下,也只有少数免疫细胞进入精小管。因此,在本研究中,我们通过巨噬细胞在体外通过SCs(含或不含小管周围细胞(PCs)、含或不含基质细胞(MG))的转移来评估睾丸免疫屏障(TIB)。原代PCs是从成年大鼠睾丸中分离出来的,并与我们小组最近获得的大鼠有条件重编程的原代成年支持细胞系(PASC1)单独或共培养。从新鲜血液中分离的大鼠单核细胞分化为M0巨噬细胞,极化后分化为M1或M2巨噬细胞,其特征是M1表达CXCL11和TNF-α, M2表达CCL17和CCL22。白细胞追踪器(leukotracker)标记的M0、M1和M2巨噬细胞在有MG和不含MG的PCs/SCs的单培养和共培养中迁移表明,SCs是体外TIB的主要成分,PCs或MG的贡献微不足道。此外,与M0或M1相比,M2巨噬细胞的迁移活性较低。与白细胞介素-6 (IL-6)或肿瘤坏死因子-α (TNF-α)的负作用相比,睾酮(T)治疗SCs对屏障有积极作用。新的迁移模型适用于评价巨噬细胞通过睾丸细胞屏障的迁移,可用于从免疫学角度研究睾丸屏障的完整性。
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来源期刊
Tissue Barriers
Tissue Barriers MEDICINE, RESEARCH & EXPERIMENTAL-
CiteScore
6.60
自引率
6.50%
发文量
25
期刊介绍: Tissue Barriers is the first international interdisciplinary journal that focuses on the architecture, biological roles and regulation of tissue barriers and intercellular junctions. We publish high quality peer-reviewed articles that cover a wide range of topics including structure and functions of the diverse and complex tissue barriers that occur across tissue and cell types, including the molecular composition and dynamics of polarized cell junctions and cell-cell interactions during normal homeostasis, injury and disease state. Tissue barrier formation in regenerative medicine and restoration of tissue and organ function is also of interest. Tissue Barriers publishes several categories of articles including: Original Research Papers, Short Communications, Technical Papers, Reviews, Perspectives and Commentaries, Hypothesis and Meeting Reports. Reviews and Perspectives/Commentaries will typically be invited. We also anticipate to publish special issues that are devoted to rapidly developing or controversial areas of research. Suggestions for topics are welcome. Tissue Barriers objectives: Promote interdisciplinary awareness and collaboration between researchers working with epithelial, epidermal and endothelial barriers and to build a broad and cohesive worldwide community of scientists interesting in this exciting field. Comprehend the enormous complexity of tissue barriers and map cross-talks and interactions between their different cellular and non-cellular components. Highlight the roles of tissue barrier dysfunctions in human diseases. Promote understanding and strategies for restoration of tissue barrier formation and function in regenerative medicine. Accelerate a search for pharmacological enhancers of tissue barriers as potential therapeutic agents. Understand and optimize drug delivery across epithelial and endothelial barriers.
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