Creation of bio-artificial kidney with renal cells (primary, immortalised or stem) as a model of renal transport
Lead Research Organisation:
Loughborough University
Department Name: Chemical Engineering
Abstract
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.
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.
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/P50483X/1 | 01/11/2016 | 30/04/2021 | |||
| 1814503 | Studentship | BB/P50483X/1 | 01/11/2016 | 31/01/2021 | Alexandros Englezakis |
| Description | The first key finding was the development and characterisation of Polysulphone and PVDF biocompatible materials that would allow cells to attach while keeping their morphology and function. Flat membranes of different biocompatible materials were made and characterised. The use of several coatings was further explored but have proven that some coatings could be toxic and cause cell death. A suitable material based on PVDF was selected due to its ease of manufacture, proximal tubule cell attachment properties. In addition, it requires no extra and complex coatings to accommodate cell adhesion while attached cells maintained their cellular function. Attempts were made for converting the same material into hollow fibre form in order to be used in a bioartificial kidney model but were not successful. These HFs were fragile, non-homogeneous highly permeable that may have been cause by fractions on the HF structure. Instead, commercially available hollow fibres were purchased, and different coatings were used in order to enhance cell adhesion on the material. Once optimised, hollow fibres were inserted in a bioartificial kidney module with cells injected in the fibre's lumen. In addition, these cells were exposed to shear stress by using different media flow rates and a RT-PCR panel was used to examine differences in proximal tubule markers gene expression when exposed to shear stress. Gene expression differences were observed between cells grown in static and flow conditions indicating that the bioartificial kidney model provides more physiological conditions to cells than conventional cell cultures. This further supports the hypothesis that a 3D cell culture has a more physiological phenotype compared to 2D cell cultures. The final part that is now examined, is the study of drug transport across the cellular monolayer and comparison with conventional 2D models. It has shown that drug transport is signifignatly affected by shear stress and enviroment alone, furhter supporting that the bioartificial kidney is provides a diffrent enviroment to 2D models. If this is more physiological to in vivo conditions is yet to be examined furhter. |
| Exploitation Route | The finding that PVDF based material is an easy to make and biocompatible material, could be used as a scaffold for cell for a number of regenerative applications. In addition, the use of coated hollow fibres with enhanced cell attachment properties could be used for large scale cell cultures. Finally, the model would be used on known and new drugs that have certain effect in vivo and compare its efficiency and accuracy in predicting renal clearance and DDI. This would allow having more accurate data in the ADMET properties of new drugs. This in turn would reduce the cost and time in drug development |
| Sectors | Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Description | ESAO (European society of artificial organs) winter school 2020 |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | 50 postgraduate and professors attented the ESAO winter school 2020 to have an update on tissue engineering. A poster on the project was presented to several members of the scociety and exchanged ideas. |
| Year(s) Of Engagement Activity | 2020 |
| URL | https://www.esao.org/events/winterschool/ |