Creation of bio-artificial kidney with renal cells (primary, immortalised or stem) as a model of renal transport

体外人肾模型的开发是早期药物开发的关键要求，以预测人的肾清除率并潜在地开发结构活性关系。本项目的目的是生成这样一个模型，该模型还可以通过肾脏转运蛋白确定肾脏清除率和药物相互作用（DDI）的体外至体内外推（IVIVE）。成功开发体外肾模型的关键因素是肾转运蛋白的表达和功能沿着紧密细胞单层的形成，其中顶侧和基底侧隔室可以分别进入和采样。拉夫堡大学先前已经开发了利用聚砜中空纤维的生物反应器系统。管状定向、应用流动模拟近端小管的体内环境以及生物反应器系统的结构组成已被证明可改善近端小管细胞生长和活力，然而，该平台对肾摄取和外排转运蛋白表达和功能的影响仍有待研究。因此，在本项目中，将使用人肾细胞（HK 2）确定细胞培养基组成、细胞密度、流速、聚砜纤维上的细胞培养表面和挤出方法方面的最佳条件，以生产用于肾细胞在生物反应器系统上生长的一致中空纤维。随后，将在生物反应器中检测人肾近端小管细胞（包括原代和永生化细胞），以评估摄取和外排转运蛋白的表达、功能和紧密细胞单层的形成。一旦选择了在生物反应器系统中具有最佳生长、活力和肾转运蛋白表达的细胞类型，它们将用作体外人肾模型，以确定肾清除率的IVIVE，预测肾分泌和通过肾转运蛋白的药物-药物相互作用。由该模型产生的数据也可以包括在基于生理学的药代动力学建模包中，以评价人体内DDI预测。

Development of an in vitro human renal model is a key requirement for early stage drug development to predict renal clearance in human and potentially develop structure activity relationships. The aim of this project is to generate such a model which can also enable the determination in vitro to in vivo extrapolation (IVIVE) for renal clearance and drug-drug interactions (DDIs) via renal transporters. Key factors for successful development of an in vitro renal model are expression and function of renal transporters along with formation of tight cellular monolayer where both apical and basolateral compartments that can be accessed and sampled separately. A bioreactor system utilising polysulphone hollow fibres has previously been developed at Loughborough University. Tubular orientation, application of flow to mimic the in vivo environment of the proximal tubule and the structural composition of bioreactor system have been shown to improve proximal tubule cell growth and viability, however, the effect of this platform on expression and function renal uptake and efflux transporters remains to be studied. In this project, human kidney cells (HK2) are therefore going to be used to determine the optimum conditions in terms of cell culture medium composition, cell density, flow rate, cell culture surface on the polysulphone fibres and extrusion method to produce consistent hollow fibres for renal cells growth on the bioreactor system. Subsequently, human renal proximal tubule cells including primary and immortalized cells will be tested in the bioreactor to assess the uptake and efflux transporters expression, function and formation of tight cellular monolayer. Once cell types with optimum growth, viability, and renal transporter expression in the bioreactor system are selected they will be utilised as an in vitro human renal model to determine an IVIVE for renal clearance, prediction of renal secretion and drug-drug interactions via renal transporters. The data produced by this model may also be included in physiologically based pharmacokinetic modelling packages to evaluate in vivo DDIs predictions in humans.