Ali Rizwan, Huwaizi Sarah, Alhallaj Alshaimaa, Al Subait Arwa, Barhoumi Tlili, Al Zahrani Hajar, Al Anazi Abdullah, Latif Khan Abdul, Boudjelal Mohamed
Medical Research Core Facility and Platforms, King Abdullah International Medical Research Center (KAIMRC), King Saud bin Abdulaziz University for Health Sciences (KSAU-HS), MNGHA, Riyadh, Saudi Arabia.
Department of Pathology and Laboratory Medicine, King Abdulaziz Medical City (KAMC), MNGHA, Riyadh, Saudi Arabia.
Front Mol Biosci. 2021 Dec 24;8:769030. doi: 10.3389/fmolb.2021.769030. eCollection 2021.
Three-dimensional (3D) cell culture systems have become very popular in the field of drug screening and discovery. There is an immense demand for highly efficient and easy methods to produce 3D spheroids in any cell format. We have developed a novel and easy method to produce spheroids from the newly isolated KAIMRC1 cell line . It can be used as a 3D model to study proliferation, differentiation, cell death, and drug response of cancer cells. Our procedure requires growth media supplemented with 10% new born calf serum (NBCS) and regular cell culture plates to generate KAIMRC1 spheroids without the need for any specialized 3D cell culture system. This procedure generates multiple spheroids within a 12-24-h culture. KAIMRC1 spheroids are compact, homogeneous in size and morphology with a mean size of 55.8 µm (±3.5). High content imaging (HCI) of KAIMRC1 spheroids treated with a panel of 240 compounds resulted in the identification of several highly specific compounds towards spheroids. Immunophenotyping of KAIMRC1 spheroids revealed phosphorylation of FAK, cJUN, and E-cadherin, which suggests the involvement of JNK/JUN pathway in the KAIMRC1 spheroids formation. Gene expression analysis showed upregulation of cell junction genes, GJB3, DSC1, CLDN5, CLDN8, and PLAU. Furthermore, co-culture of KAIMRC1 cells with primary cancer-associated-fibroblasts (CAFs) showcased the potential of these cells in drug discovery application.
三维（3D）细胞培养系统在药物筛选和发现领域已变得非常流行。对于以任何细胞形式高效且简便地生产3D球体的方法有着巨大需求。我们开发了一种新颖且简便的方法，可从新分离的KAIMRC1细胞系中生成球体。它可作为一种3D模型，用于研究癌细胞的增殖、分化、细胞死亡及药物反应。我们的方法需要添加10%新生小牛血清（NBCS）的生长培养基和常规细胞培养板来生成KAIMRC1球体，无需任何专门的3D细胞培养系统。此方法在12 - 24小时培养内可生成多个球体。KAIMRC1球体紧密，大小和形态均一，平均大小为55.8 µm（±3.5）。用一组240种化合物处理KAIMRC1球体的高内涵成像（HCI）结果鉴定出了几种对球体具有高度特异性的化合物。KAIMRC1球体的免疫表型分析显示了粘着斑激酶（FAK）、cJUN和E - 钙粘蛋白的磷酸化，这表明JNK/JUN通路参与了KAIMRC1球体的形成。基因表达分析显示细胞连接基因GJB3、DSC1、CLDN5、CLDN8和PLAU上调。此外，KAIMRC1细胞与原发性癌症相关成纤维细胞（CAF）的共培养展示了这些细胞在药物发现应用中的潜力。
