Allcock Benjamin, Wei Wenbin, Goncalves Kirsty, Hoyle Henry, Robert Alisha, Quelch-Cliffe Rebecca, Hayward Adam, Cooper Jim, Przyborski Stefan
Department of Biosciences, Durham University, Durham DH1 3LE, UK.
European Collection of Authenticated Cell Cultures, Salisbury SP4 0JG, UK.
Cells. 2023 Oct 5;12(19):2408. doi: 10.3390/cells12192408.
It is widely recognised that cells respond to their microenvironment, which has implications for cell culture practices. Growth cues provided by 2D cell culture substrates are far removed from native 3D tissue structure in vivo. Geometry is one of many factors that differs between in vitro culture and in vivo cellular environments. Cultured cells are far removed from their native counterparts and lose some of their predictive capability and reliability. In this study, we examine the cellular processes that occur when a cell is cultured on 2D or 3D surfaces for a short period of 8 days prior to its use in functional assays, which we term: "priming". We follow the process of mechanotransduction from cytoskeletal alterations, to changes to nuclear structure, leading to alterations in gene expression, protein expression and improved functional capabilities. In this study, we utilise HepG2 cells as a hepatocyte model cell line, due to their robustness for drug toxicity screening. Here, we demonstrate enhanced functionality and improved drug toxicity profiles that better reflect the in vivo clinical response. However, findings more broadly reflect in vitro cell culture practises across many areas of cell biology, demonstrating the fundamental impact of mechanotransduction in bioengineering and cell biology.
人们普遍认识到，细胞会对其微环境做出反应，这对细胞培养实践具有重要意义。二维细胞培养底物提供的生长信号与体内天然的三维组织结构相差甚远。几何形状是体外培养和体内细胞环境之间存在差异的众多因素之一。培养的细胞与其天然对应物有很大不同，并且失去了一些预测能力和可靠性。在本研究中，我们研究了在用于功能测定之前，将细胞在二维或三维表面上培养8天这一短时间内发生的细胞过程，我们将其称为“预刺激”。我们追踪从细胞骨架改变到核结构变化，再到基因表达、蛋白质表达改变以及功能能力改善的机械转导过程。在本研究中，我们使用HepG2细胞作为肝细胞模型细胞系，因为它们在药物毒性筛选方面具有较强的耐受性。在此，我们展示了增强的功能和改善的药物毒性概况，能更好地反映体内临床反应。然而，这些发现更广泛地反映了细胞生物学许多领域的体外细胞培养实践，证明了机械转导在生物工程和细胞生物学中的根本影响。
