This project has two aims; (1) to replace animals in an important field of research and (2) to develop a superior culture system for renal tissue engineering. Research into renal regeneration and renal replacement therapy is growing rapidly, driven by urgent clinical need. It is clear, however, that advance will depend on a better understanding of signalling and lineage in normal renal development. These are being investigated in a variety of ways, all of which use animals, either in vivo or by organ culture of kidneys isolated from embryos. The organ culture system has proved particularly useful, but is still limited in its ability to allow manipulation of gene expression. This project aims to produce cell lines that can give rise to specific components of the kidney in an organotypic setting, and can reconstitute a whole embryonic kidney in combination. Based on our existing systems, we will produce these cell lines both by conditional immortalization of cells from embryonic kidneys and by sorting and culture of ES cell derivatives. We will assess the fates available to each line in an organotypic context and will assess their ability to produce renal structures, up to a whole kidney, in novel tissue engineering culture systems based on our current state-of-the-art. This project brings together the expertise of three laboratories to achieve a significant advance in both the 3Rs and in renal regeneration and replacement by tissue engineering.
- Research Review 2011: Replacing the use of animals with kidney cell lines
Davies JA (2015) Biological techniques: Kidney tissue grown from induced stem cells. Nature 526(7574): 512-3 doi:10.1038/nature15639
Davies JA (2015) Self-organized Kidney Rudiments: Prospects for Better in vitro Nephrotoxicity Assays. Biomarker Insights 10(Suppl 1): 117-23 doi:10.4137/BMI.S20056
Tai G, Hohenstein P, Davies JA (2013) FAK-Src signalling is important to renal collecting duct morphogenesis: discovery using a hierarchical screening technique. Biol Open 2(4):416-23 doi:10.1242/bio.20133780
Davies JA (2012) Fetal Organ Culture. Replacing Animal Models: A Practical Guide to Creating and Using Culture-Based Biomimetic Alternatives 81-87 doi: 10.1002/9781119940685.ch8
Davies JA (2012) Potential Advantages of Using Biomimetic Alternatives. Replacing Animal Models: A Practical Guide to Creating and Using Culture-Based Biomimetic Alternatives 1-11 doi:10.1002/9781119940685.ch1
Davies JA, Unbekandt M, Ineson J, Lusis M, Little MH (2012) Dissociation of embryonic kidney followed by re-aggregation as a method for chimeric analysis.Methods Mol Biol 886:135-46 doi:10.1007/978-1-61779-851-1_12.
Chang CH and Davies JA (2012) An improved method of renal tissue engineering, by combining renal dissociation and reaggregation with a low-volume culture technique, results in development of engineered kidneys complete with loops of Henle. Nephron Exp Nephrol. 121(3-4):e79-85. doi:10.1159/000345514
Tai G and Davies JA (2012) Hierarchical Screening of Pathways: Using Cell and Organ Cultures to Reduce use of Transgenic Mice. Replacing Animal Models: A Practical Guide to Creating and Using Culture-Based Biomimetic Alternatives 123-136 doi: 10.1002/9781119940685.ch12
Ganeva V, Unbekandt M, Davies JA (2011) An improved kidney dissociation and reaggregation culture system results in nephrons arranged organotypically around a single duct system. Organogenesis 7(2):83-7. doi:10.4161/org.7.2.14881.
Lee WC, Hough MT, Liu W, Ekiert R, Lindström NO, Hohenstein P, Davies JA (2010) Dact2 is expressed in the developing ureteric bud/collecting duct system of the kidney and controls morphogenetic behavior of collecting duct cells. Am J Physiol Renal Physiol 299(4):F740-51. doi:10.1152/ajprenal.00148.2010.
Sebinger DD, Unbekandt M, Ganeva VV, Ofenbauer A, Werner C, Davies JA (2010) A novel, low-volume method for organ culture of embryonic kidneys that allows development of cortico-medullary anatomical organization.PLoS One 5(5):e10550. doi:10.1371/journal.pone.0010550.