Microfluidic proximal tubule kidney-on-chip
The present invention relates to microfluidic fluidic devices, methods and systems as microfluidic kidney on-chips, e.g. human Proximal Tubule-Kidney-Chip, Glomerulus (Kidney)-Chip, Collecting Duct (Kidney)-Chip. Devices, methods and systems are described for drug testing including drug transport and renal clearance. Further, such devices, methods and systems are used for determining drug-drug interactions and their effect upon renal transporter functions. Importantly, they may be used for pre-clinical and clinical drug development for treating kidney diseases and for personalized medicine.
1. A method of measuring active transport, comprising:
a) providing a microfluidic device comprising a membrane, said membrane comprising a first surface and a second surface, said first surface comprising proximal tubule cells and said second surface comprising glomerular microvascular endothelial cells, wherein said first surface of said membrane is part of a first microfluidic channel and said second surface of said membrane is part of a second microfluidic channel;
b) culturing said proximal tubule cells and said glomerular microvascular endothelial cells with media under continuous low shear flow;
c) introducing an agent; and
d) detecting active transport of said agent, comprising greater transport of said agent, from said second microfluidic channel to said first microfluidic channel as compared to transport of said agent from said first microfluidic channel to said second microfluidic channel.
2. The method of claim 1 , wherein said proximal tubule cells are human primary proximal tubular epithelial cells.
3. The method of claim 2 , wherein said membrane contains pores.
4. The method of claim 1 , wherein said transport detected in step d) is in the presence of one or more inhibitors.
5. The method of claim 1 , wherein said transport detected in step d) comprises a measurement in the effluent.
6. The method of claim 1 , wherein said agent introduced in step c) is a therapeutic drug.
7. The method of claim 1 , wherein said agent introduced in step c) is metformin.
8. The method of claim 1 , wherein there is an approximately 2-fold increase in net transport of said agent from said second channel to said first channel compared to net transport from said first channel to said second channel.
9. The method of claim 1 , wherein said agent introduced in step c) is creatinine.
10. The method of claim 1 , wherein said agent introduced in step c) is p-aminohippuric acid.
11. The method of claim 9 , wherein said transport of said creatinine from said second microfluidic channel to said first microfluidic channel is approximately 2-fold greater as compared to the transport of said creatinine from said first microfluidic channel to said second microfluidic channel.
12. The method of claim 10 , wherein said transport of said p-aminohippuric acid from said second microfluidic channel to said first microfluidic channel is approximately 2-fold greater as compared to the transport of said p-aminohippuric acid from said first microfluidic channel to said second microfluidic channel.