Abstract:
Fibroblastic reticular cells (FRCs) are mesenchymal stromal cells in human lymph nodes (LNs) playing a pivotal role in adaptive immunity. Several FRC subsets have been identified, yet it remains to be elucidated if their heterogeneity is maintained upon culture. Here, we established a protocol to preserve and culture FRCs from human LNs and characterized their phenotypic profile in fresh LN suspensions and upon culture using multispectral flow cytometry. We found nine FRC subsets in fresh human LNs, independent of donor, of which four persisted in culture throughout several passages. Interestingly, the historically FRC-defining marker podoplanin (PDPN) was not present on all FRC subsets. Therefore, we propose that CD45negCD31neg human FRCs are not restricted by PDPN expression, as we found CD90, BST1, and CD146/MCAM to be more widely expressed. Together, our data provide insight into FRC heterogeneity in human LNs, enabling further investigation into the function of individual FRC subsets.
摘要:
成纤维细胞网状细胞(FRC)是人淋巴结(LN)中的间充质基质细胞,在适应性免疫中起关键作用。已经确定了几个FRC子集,然而,如果它们的异质性在培养时保持不变,还有待阐明。这里,我们建立了一个方案来保存和培养来自人类LN的FRC,并使用多光谱流式细胞术在新鲜的LN悬浮液中和培养时表征了它们的表型特征。我们在新鲜的人类LN中发现了9个FRC子集,独立于捐赠者,其中四个在几个段落中坚持文化。有趣的是,历史上定义FRC的标记后planin(PDPN)并未出现在所有FRC亚群上.因此,我们认为CD45negCD31neg人FRC不受PDPN表达的限制,因为我们发现CD90,BST1和CD146/MCAM更广泛地表达。一起,我们的数据提供了对人类LN中FRC异质性的洞察,能够进一步调查各个FRC子集的功能。