Université de Limoges, UMR INSERM 1308 CAPTuR, Faculté de Médecine, 87025 Limoges, France; Université de Bordeaux, INSERM U1212, UMR CNRS 5320, F-33076 Bordeaux, France.
Université de Bordeaux, INSERM U1212, UMR CNRS 5320, F-33076 Bordeaux, France; Université de Pau et des Pays de l'Adour (E2S/UPPA) CNRS, IPREM, UMR 5254, 64053 Pau Cedex, France.
J Chromatogr A. 2024 Nov 8;1736:465393. doi: 10.1016/j.chroma.2024.465393. Epub 2024 Sep 22.
Three-dimensional (3D) cancer models, such as multicellular tumor spheroids (MCTS), are biological supports used for research in oncology, drug development and nanotoxicity assays. However, due to various analytical and biological challenges, the main recurring problem faced when developing this type of 3D model is the lack of reproducibility. When using a 3D support to assess the effect of biologics, small molecules or nanoparticles, it is essential that the support remains constant over time and multiples productions. This constancy ensures that any effect observed following molecule exposure can be attributed to the molecule itself and not to the heterogeneous properties of the 3D support. In this study, we address these analytical challenges by evaluating for the first time the 3D culture of a sub-population of cancer stem cells (CSCs) from a glioblastoma cancer cell line (U87-MG), produced by a SdFFF (sedimentation field-flow fractionation) cell sorting, in a supramolecular hydrogel composed of single, well-defined molecule (bis-amide bola amphiphile 0.25% w/v) with a stiffness of 0.4 kPa. CSCs were chosen for their ability of self-renewal and multipotency that allow them to generate fully-grown tumors from a small number of cells. The results demonstrate that CSCs cultured in the hydrogel formed spheroids with a mean diameter of 336.67 ± 38.70 µm by Day 35, indicating reproducible growth kinetics. This uniformity is in contrast with spheroids derived from unsorted cells, which displayed a more heterogeneous growth pattern, with a mean diameter of 203.20 ± 102.93 µm by Day 35. Statistical analysis using an unpaired t-test with unequal variances confirmed that this difference in spheroid size is significant, with a p-value of 0.0417 (p < 0.05). These findings demonstrate that CSC-derived spheroids, when cultured in a well-defined hydrogel, exhibit highly reproducible growth patterns compared to spheroids derived from unsorted cells, making them a more reliable 3D model for biological research and drug testing applications.
三维（3D）癌症模型，如多细胞肿瘤球体（MCTS），是用于肿瘤学研究、药物开发和纳米毒性测定的生物支架。然而，由于各种分析和生物学挑战，开发这种 3D 模型时主要面临的问题是缺乏可重复性。当使用 3D 支架来评估生物制剂、小分子或纳米颗粒的效果时，支架必须随着时间和多次生产保持稳定。这种稳定性确保了在暴露于分子后观察到的任何效果都归因于分子本身，而不是 3D 支架的异质特性。在这项研究中，我们通过首次评估由沉降场流分离（sedimentation field-flow fractionation，SdFFF）细胞分选产生的神经胶质瘤癌细胞系（U87-MG）的亚群癌细胞干细胞（CSCs）在由单个明确定义的分子（双酰胺 bola 两亲物 0.25%w/v）组成的超分子水凝胶中的 3D 培养来解决这些分析挑战，该超分子水凝胶的硬度为 0.4kPa。选择 CSCs 是因为它们具有自我更新和多能性的能力，使它们能够从少量细胞中产生完全成熟的肿瘤。结果表明，在水凝胶中培养的 CSCs 在第 35 天形成了平均直径为 336.67 ± 38.70 µm 的球体，表明可重复的生长动力学。这种均匀性与未分选细胞衍生的球体形成鲜明对比，后者在第 35 天的平均直径为 203.20 ± 102.93 µm，显示出更异质的生长模式。使用具有不等方差的未配对 t 检验的统计分析证实，这种球体大小的差异是显著的，p 值为 0.0417（p < 0.05）。这些发现表明，与未分选细胞衍生的球体相比，当在明确定义的水凝胶中培养时，CSC 衍生的球体表现出高度可重复的生长模式，使它们成为更可靠的用于生物学研究和药物测试应用的 3D 模型。
