Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Bucharest, 91-95 Splaiul Independentei, Bucharest, R-050095, Romania.
Cell Biochem Biophys. 2022 Mar;80(1):63-73. doi: 10.1007/s12013-021-00982-1. Epub 2021 Apr 26.
Hanging drop represents a simple approach designed for the generation of 3D models that have potential to be used for the study of solid tumors characteristics. The aim of the study was to develop and characterize the breast cancer 3D cellular models obtained through hanging drop technique using MDA-MB-231 cells. The biological characteristics such as: morphology, cellular viability, proliferation capacity and hypoxia, were monitored for a six-day time period. The morphological evaluation indicated that the 3D models presented the aspect of compact (seeding density of 2500 and 5000 cells/drop) and loose (seeding density of 8000 cells/drop) aggregates, with a decrease in diameter and an increase of their circularity. The cellular viability and proliferation capacity decreased in time and the level of lactate dehydrogenase (LDH) increased in a time-dependent manner, suggesting the presence of necrotic cells that were dispersed in the cellular aggregates. The occurrence of hypoxia process was suggested by the up-regulation of Hsp70 protein expression and increased level of nitric oxide (NO). Moreover, the up-regulation of HIF-1α and poli-ubiquitinated Nrf2 protein expressions and decreased level of reduced glutathione (GSH) indicated the presence of an acute hypoxic environment in MDA-MB-231 3D aggregates. In conclusion, the MDA-MB-231 3D models generated through hanging drop are compact and loose aggregates characterized by an acute hypoxic condition.
悬滴法是一种简单的方法，旨在生成可用于研究实体瘤特征的 3D 模型。本研究的目的是使用 MDA-MB-231 细胞通过悬滴技术开发和表征乳腺癌 3D 细胞模型。在六天的时间内监测了生物学特性，如形态、细胞活力、增殖能力和缺氧。形态评估表明，3D 模型呈现出致密（每滴接种密度为 2500 和 5000 个细胞）和疏松（每滴接种密度为 8000 个细胞）聚集物的形态，直径减小，圆形度增加。细胞活力和增殖能力随时间降低，乳酸脱氢酶（LDH）水平呈时间依赖性增加，表明存在分散在细胞聚集体中的坏死细胞。热休克蛋白 70（Hsp70）蛋白表达上调和一氧化氮（NO）水平增加提示缺氧过程的发生。此外，缺氧诱导因子 1α（HIF-1α）和多泛素化核因子红细胞 2（Nrf2）蛋白表达上调和还原型谷胱甘肽（GSH）水平降低表明 MDA-MB-231 3D 聚集体中存在急性缺氧环境。总之，通过悬滴法生成的 MDA-MB-231 3D 模型是具有急性缺氧条件的致密和疏松聚集物。
