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经肺动脉导管获取的人肺动脉内皮细胞的体外原代培养的渐进经验。

Incremental Experience in In Vitro Primary Culture of Human Pulmonary Arterial Endothelial Cells Harvested from Pulmonary Arterial Catheters.

机构信息

Laboratory of Respiratory Diseases & Thoracic Surgery (BREATHE), Department of Chronic Diseases & Metabolism (CHROMETA) & Biomedical MRI, Department of Imaging and Pathology, University of Leuven, 3000 Leuven, Belgium.

Laboratory of Respiratory Diseases & Thoracic Surgery (BREATHE), Department of Chronic Diseases & Metabolism (CHROMETA), University of Leuven, 3000 Leuven, Belgium.

出版信息

Cells. 2021 Nov 19;10(11):3229. doi: 10.3390/cells10113229.

Abstract

Pulmonary arterial hypertension (PAH) is a devastating condition affecting the pulmonary microvascular wall and endothelium, resulting in their partial or total obstruction. Despite a combination of expensive vasodilatory therapies, mortality remains high. Personalized therapeutic approaches, based on access to patient material to unravel patient specificities, could move the field forward. An innovative technique involving harvesting pulmonary arterial endothelial cells (PAECs) at the time of diagnosis was recently described. The aim of the present study was to fine-tune the initial technique and to phenotype the evolution of PAECs in vitro subcultures. PAECs were harvested from pulmonary arterial catheters during routine diagnostic or follow up right heart catheterization. Collected PAECs were phenotyped by flow cytometry and immunofluorescence focusing on endothelial-specific markers. We highlight the ability to harvest patients' PAECs and to maintain them for up to 7-12 subcultures. By tracking the endothelial phenotype, we observed that PAECs could maintain an endothelial phenotype for several weeks in culture. The present study highlights the unique opportunity to obtain homogeneous subcultures of primary PAECs from patients at diagnosis and follow-up. In addition, it opens promising perspectives regarding tailored precision medicine for patients suffering from rare pulmonary vascular diseases.

摘要

肺动脉高压(PAH)是一种严重的疾病,影响肺微血管壁和内皮细胞,导致它们部分或完全阻塞。尽管采用了昂贵的血管扩张疗法的联合治疗,死亡率仍然很高。基于获取患者材料以揭示患者特异性的个性化治疗方法可能会推动该领域的发展。最近描述了一种涉及在诊断时采集肺动脉内皮细胞(PAEC)的创新技术。本研究的目的是微调初始技术,并对体外亚培养物中 PAEC 的表型进行研究。从常规诊断或随访右心导管检查期间的肺动脉导管中采集 PAEC。通过流式细胞术和免疫荧光法对收集的 PAEC 进行表型分析,重点关注内皮特异性标志物。我们强调了从患者中采集 PAEC 并将其维持长达 7-12 代的能力。通过跟踪内皮表型,我们观察到 PAEC 在培养物中可以维持数周的内皮表型。本研究突出了从诊断和随访时的患者中获得同质的原发性 PAEC 亚培养物的独特机会。此外,它为患有罕见肺血管疾病的患者提供了针对精准医学的有前景的前景。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ed8/8618201/fd1a9725e5e0/cells-10-03229-g001.jpg

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