Wu Cheng-Guang, Chiovaro Francesca, Curioni-Fontecedro Alessandra, Casanova Ruben, Soltermann Alex
Institute of Pathology and Molecular Pathology, University Hospital Zurich, Schmelzbergstrasse 12, 8091, Zurich, Switzerland.
University of Zurich, Rämistrasse 71, 8006, Zurich, Switzerland.
J Transl Med. 2020 Apr 10;18(1):163. doi: 10.1186/s12967-020-02331-x.
Malignant serous effusion (MSE) denotes a manifestation of metastatic disease with typical high concentrations of both cancer and immune cells, making them an ideal resource for in vitro cytologic studies. Hence, the aim of the study was to investigate the features of 2D and 3D MSE culture systems as well as their feasibilities for in vitro drug screening.
Pleural and peritoneal effusions from 8 patients were collected and processed for 2D monolayer and 3D hanging drop cell culture into GravityPLUS™ plates. Representative markers for cell components, proliferation rate and tumour classification were investigated by immunohistochemistry, followed by absolute quantification using a digitalised image analysis approach. Further, we implemented another 3D cell culture model based on a low attachment method for in vitro drug sensitivity testing of carboplatin, pemetrexed and pembrolizumab for 5 patients.
Monolayer cell culture was favourable for the growth of mesothelial cells, while hanging drop culture in GravityPLUS™ plates showed better ability for preserving cancer cells, inducing positive diagnostic markers expression and restraining the growth of mesothelial cells. For in vitro drug testing, MSE from five patients presented various drug sensitivities, and one case showed strong response to PD-1 checkpoint inhibition (pembrolizumab). For some patients, the application of combinatorial drugs had better therapeutic responses compared to monotherapy.
Digitalised quantification of data offers a better understanding of different MSE culture models. More importantly, the proposed platforms are practical and amenable for performing in vitro chemo-/immunotherapeutic drug testing by using routine cytologic MSE in a personalised manner. Next to cell blocks, our work demonstrates the prognostic and predictive value of cytologic effusion samples.
恶性浆液性积液（MSE）是转移性疾病的一种表现，其中癌细胞和免疫细胞浓度通常较高，使其成为体外细胞学研究的理想资源。因此，本研究的目的是探讨二维和三维MSE培养系统的特征及其在体外药物筛选中的可行性。
收集8例患者的胸腔和腹腔积液，处理后用于二维单层和三维悬滴细胞培养，接种到GravityPLUS™平板中。通过免疫组织化学研究细胞成分、增殖率和肿瘤分类的代表性标志物，随后使用数字化图像分析方法进行绝对定量。此外，我们基于低附着方法实施了另一种三维细胞培养模型，用于对5例患者的卡铂、培美曲塞和帕博利珠单抗进行体外药敏试验。
单层细胞培养有利于间皮细胞生长，而在GravityPLUS™平板中进行悬滴培养在保存癌细胞、诱导阳性诊断标志物表达和抑制间皮细胞生长方面表现出更好的能力。对于体外药物测试，5例患者的MSE呈现出不同的药物敏感性，1例对PD-1检查点抑制（帕博利珠单抗）表现出强烈反应。对于一些患者，联合用药比单一疗法具有更好的治疗反应。
数据的数字化定量有助于更好地理解不同的MSE培养模型。更重要的是，所提出的平台实用且适合以个性化方式使用常规细胞学MSE进行体外化学/免疫治疗药物测试。除了细胞块，我们的工作证明了细胞学积液样本的预后和预测价值。
