Oliveira Fabiana, Bondareva Olga, Rodríguez-Aguilera Jesús Rafael, Sheikh Bilal N
Helmholtz Institute for Metabolic, Obesity and Vascular Research (HI-MAG) of the Helmholtz Center Munich, Leipzig, Germany.
Medical Faculty, Leipzig University, Leipzig, Germany.
Front Cell Neurosci. 2023 May 2;17:1165887. doi: 10.3389/fncel.2023.1165887. eCollection 2023.
Pericytes (PCs) are essential components of the blood brain barrier. Brain PCs are critical for dynamically regulating blood flow, for maintaining vascular integrity and their dysregulation is associated with a myriad of disorders such as Alzheimer's disease. To understand their physiological and molecular functions, studies have increasingly focused on primary brain PC isolation and culture. Multiple methods for PC culture have been developed over the years, however, it is still unclear how primary PCs compare to their counterparts. To address this question, we compared cultured brain PCs at passage 5 and 20 to adult and embryonic brain PCs directly isolated from mouse brains via single cell RNA-seq. Cultured PCs were highly homogeneous, and were most similar to embryonic PCs, while displaying a significantly different transcriptional profile to adult brain PCs. Cultured PCs downregulated canonical PC markers and extracellular matrix (ECM) genes. Importantly, expression of PC markers and ECM genes could be improved by co-culture with brain endothelial cells, showing the importance of the endothelium in maintaining PC identity and function. Taken together, these results highlight key transcriptional differences between cultured and PCs which should be considered when performing experiments with brain PCs.
周细胞（PCs）是血脑屏障的重要组成部分。脑周细胞对于动态调节血流、维持血管完整性至关重要，其功能失调与多种疾病相关，如阿尔茨海默病。为了解其生理和分子功能，研究越来越多地聚焦于原代脑周细胞的分离和培养。多年来已开发出多种周细胞培养方法，然而，原代周细胞与其他周细胞相比情况仍不清楚。为解决这个问题，我们通过单细胞RNA测序，将第5代和第20代培养的脑周细胞与直接从小鼠脑部分离的成年和胚胎脑周细胞进行了比较。培养的周细胞高度均一，与胚胎周细胞最为相似，而与成年脑周细胞的转录谱有显著差异。培养的周细胞下调了典型的周细胞标志物和细胞外基质（ECM）基因。重要的是，通过与脑内皮细胞共培养可以改善周细胞标志物和ECM基因的表达，这表明内皮细胞在维持周细胞特性和功能方面的重要性。综上所述，这些结果突出了培养的周细胞与其他周细胞之间的关键转录差异，在进行脑周细胞实验时应予以考虑。
