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

体外人类肾脏模型的开发是早期药物开发的关键要求，以预测人类肾脏清除并潜在地发展结构活性关系。该项目的目的是建立这样一个模型，该模型也可以通过肾脏转运蛋白测定肾脏清除率和药物-药物相互作用（ddi）的体外-体内外推法（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，以及紧密细胞单层的形成，其中顶端和基底外侧的室室都可以单独进入和采样。拉夫堡大学以前开发了一种利用聚砜中空纤维的生物反应器系统。小管定向、应用流体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以改善近端小管细胞的生长和活力，然而，该平台对肾摄取和外排转运蛋白的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（HK2）来确定最佳条件，包括细胞培养基成分、细胞密度、流速、聚砜纤维上的细胞培养表面和挤出方法，以产生一致的中空纤维，供肾细胞在生物反应器系统上生长。随后，将在生物反应器中测试人肾近端小管细胞，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。一旦在生物反应器系统中选择了具有最佳生长、活力和肾转运蛋白表达的细胞类型，它们将被用作体外人肾模型，以确定通过肾转运蛋白进行肾脏清除、预测肾脏分泌和药物-药物相互作用的活体活体细胞（IVIVE）。该模型产生的数据也可以包含在基于生理学的药代动力学建模包中，以评估人类体内ddi的预测。

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.