Ncube Keith N, Jurgens Tamarin, Steenkamp Vanessa, Cromarty Allan D, van den Bout Iman, Cordier Werner
Department of Pharmacology, Faculty of Health Sciences, University of Pretoria, Pretoria 0007, South Africa.
Department of Physiology, Faculty of Health Sciences, University of Pretoria, Pretoria 0007, South Africa.
Biomedicines. 2023 May 19;11(5):1484. doi: 10.3390/biomedicines11051484.
Three-dimensional cell culture models are increasingly adopted as preferred pre-clinical drug testing platforms, as they circumvent limitations associated with traditional monolayer cell cultures. However, many of these models are not fully characterized. This study aimed to characterize a BT-20 triple-negative breast carcinoma spheroid model and assess its susceptibility to doxorubicin in comparison to a monolayer model. Spheroids were developed using the liquid overlay method. Phenotypic attributes were analyzed by characterizing changes in size, gross morphology, protein content, metabolic activity, hypoxic status, and cell-cell junctions. The cytotoxic range of doxorubicin in monolayers was determined using the sulforhodamine B assay, and the comparative effect of toxic and sub-toxic concentrations was assessed in both spheroids and monolayers. Similar to the in vivo microenvironment, spheroids had a heterogeneous spatial cytoarchitecture, inherent hypoxia and strong adherens junctions. Doxorubicin induced dose-dependent cytotoxicity in monolayers (IC: 130 nM, IC: 320 nM and IC: 1580 nM); however, these concentrations did not alter the spheroid size or acid phosphatase activity. Only concentrations ≥6 µM had any effect on spheroid integrity. In comparison to monolayers, the BT-20 spheroid model has decreased sensitivity to doxorubicin and could serve as a better model for susceptibility testing in triple-negative breast cancer.
三维细胞培养模型越来越多地被用作首选的临床前药物测试平台，因为它们规避了与传统单层细胞培养相关的局限性。然而，这些模型中的许多并未得到充分表征。本研究旨在表征一种BT-20三阴性乳腺癌球体模型，并与单层模型相比评估其对多柔比星的敏感性。球体采用液体覆盖法构建。通过表征大小、总体形态、蛋白质含量、代谢活性、缺氧状态和细胞间连接的变化来分析表型特征。使用磺酰罗丹明B测定法确定多柔比星在单层中的细胞毒性范围，并在球体和单层中评估毒性和亚毒性浓度的比较效果。与体内微环境相似，球体具有异质的空间细胞结构、内在缺氧和强黏附连接。多柔比星在单层中诱导剂量依赖性细胞毒性（IC：130 nM，IC：320 nM和IC：1580 nM）；然而，这些浓度并未改变球体大小或酸性磷酸酶活性。仅浓度≥6 µM对球体完整性有任何影响。与单层相比，BT-20球体模型对多柔比星的敏感性降低，可作为三阴性乳腺癌敏感性测试的更好模型。
