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Normal mesothelial cell lines newly derived from human pleural biopsy explants.
American Journal of Physiology-Lung Cellular and Molecular Physiology ( IF 3.6 ) Pub Date : 2020-07-29 , DOI: 10.1152/ajplung.00141.2020
Nathanael Pruett 1 , Anand Singh 1 , Ahjeetha Shankar 1 , David S Schrump 1 , Chuong D Hoang 1
Affiliation  

Mesothelial cells are arranged as a monolayer on covering membranes that invest surfaces of body cavities like the pleura and peritoneum. Primary human mesothelial cell (HMC) cultures are needed for studying mesothelial cell homeostasis and developing disease models, such as wound healing or cancers. Remarkably, there is a paucity of useable HMC lines that are currently available which faithfully recapitulate normal, in vivo phenotypic characteristics. Here, we present a strategy to recover HMC from human pleural tissue and to immortalize them for extended in vitro culturing. Human pleural membrane was harvested by minimally invasive surgical techniques. HMC were isolated using a two-step process combining explant cellular outgrowth from biopsy tissue and flow cytometry based on cell surface expression of cadherin-1 and CD71. Cell cultures were generated after lentiviral transfection with human telomerase. The new HMC cultures retain the same phenotypic traits and physiologic features as their in vivo counterparts, yet they can be adapted for short-term or long-term culture in large-scale in vitro experimentation. In particular, we generated a new HMC line harboring a germline mutation in BAP1, a causal tumor suppressor gene, that could be instrumental to malignant mesothelioma research. Patient-specific, normal HMC may serve as novel discovery tools allowing more powerful research models of both normal physiology and disease processes. Our surgically driven approach leads to a limitless resource of novel mesothelial cell cultures.

中文翻译:

新衍生自人胸膜活检外植体的正常间皮细胞系。

间皮细胞在覆盖膜上排列成单层,覆盖体腔表面,如胸膜和腹膜。研究间皮细胞稳态和开发疾病模型(如伤口愈合或癌症)需要原代人类间皮细胞 (HMC) 培养物。值得注意的是,目前可用的可用 HMC 系很少,它们忠实地概括了正常的体内表型特征。在这里,我们提出了一种从人类胸膜组织中恢复 HMC 并使其永生化以进行体外培养的策略。人类胸膜是通过微创手术技术获得的。使用两步法分离 HMC,结合活检组织的外植体细胞生长和基于 cadherin-1 和 CD71 的细胞表面表达的流式细胞术。用人端粒酶慢病毒转染后产生细胞培养物。新的 HMC 培养物保留了与其体内对应物相同的表型特征和生理特征,但它们可以适应大规模体外实验中的短期或长期培养。特别是,我们生成了一个新的 HMC 系,它在 BAP1(一种因果肿瘤抑制基因)中含有种系突变,这可能有助于恶性间皮瘤的研究。特定于患者的正常 HMC 可以作为新的发现工具,允许对正常生理学和疾病过程进行更强大的研究模型。我们的手术驱动方法导致了新的间皮细胞培养物的无限资源。新的 HMC 培养物保留了与其体内对应物相同的表型特征和生理特征,但它们可以适应大规模体外实验中的短期或长期培养。特别是,我们生成了一个新的 HMC 系,它在 BAP1(一种因果肿瘤抑制基因)中含有种系突变,这可能有助于恶性间皮瘤的研究。特定于患者的正常 HMC 可以作为新的发现工具,允许对正常生理学和疾病过程进行更强大的研究模型。我们的手术驱动方法导致了新的间皮细胞培养物的无限资源。新的 HMC 培养物保留了与其体内对应物相同的表型特征和生理特征,但它们可以适应大规模体外实验中的短期或长期培养。特别是,我们生成了一个新的 HMC 系,它在 BAP1(一种因果肿瘤抑制基因)中含有种系突变,这可能有助于恶性间皮瘤的研究。特定于患者的正常 HMC 可以作为新的发现工具,允许对正常生理学和疾病过程进行更强大的研究模型。我们的手术驱动方法导致了新的间皮细胞培养物的无限资源。一种因果肿瘤抑制基因,可能有助于恶性间皮瘤的研究。特定于患者的正常 HMC 可以作为新的发现工具,允许对正常生理学和疾病过程进行更强大的研究模型。我们的手术驱动方法导致了新的间皮细胞培养物的无限资源。一种因果肿瘤抑制基因,可能有助于恶性间皮瘤的研究。特定于患者的正常 HMC 可以作为新的发现工具,允许对正常生理学和疾病过程进行更强大的研究模型。我们的手术驱动方法导致了新的间皮细胞培养物的无限资源。
更新日期:2020-08-20
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