A Novel, Low-Volume Method for Organ Culture of Embryonic Kidneys That Allows Development of Cortico-Medullary Anatomical Organization

Sebinger, David Daniel Raphael; Unbekandt, Mathieu; Ganeva, Veronika V.; Ofenbauer, Andreas; Werner, Carsten; Davies, Jamie A.

doi:10.1371/journal.pone.0010550

Cited by 57 publications

(48 citation statements)

References 48 publications

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“…Previous reports have used surface area expansion as a measurement for renal development (7,12,13). Our study shows no correlation between the surface expansion and a more direct kidney developmental marker such as ureteric bud count.…”

Section: Discussioncontrasting

confidence: 63%

“…Based on previous studies (7,12,13), a relationship between metanephric size expansion and ureteric tip count, a more direct measurement of renal development, was expected. However, as can be noted in Figure 1a, surface area expansion did not correlate with ureteric tip formation (R 2 = 0.005, P = 0.77) and was therefore not used as a marker in our studies.…”

Section: Metanephric Growthmentioning

confidence: 99%

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Antibiotics and renal branching morphogenesis: comparison of toxicities

et al. 2014

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Background: Many premature born neonates receive antibiotic drugs to treat infections, which are applied during active nephrogenesis. We studied the impact of clinical concentrations of gentamicin and alternatives, ceftazidime and meropenem, on ureteric branching. Methods: Mice metanephroi were dissected at embryonic day 13 and cultured in media with or without various concentrations of gentamicin, ceftazidime, or meropenem. Zero and 24 h kidney size were assessed by surface area measurements, and the ureteric tree was visualized by whole mount staining and confocal microscopy. Branching was evaluated by counting and gene expression levels of Wt1, Sox9, Bmp7, Fgf8, and Gdnf were investigated. results: A concentration of 2,000 μmol/l ceftazidime impaired ureteric development. In addition, a 4.5-fold and a 2.5-fold downregulation was noted in Fgf8 and Gdnf, respectively. No adverse effects were noted after gentamicin or meropenem treatment. No relationship was noted between surface area expansion and ureteric bud formation, but surface area at explantation related to bud count after 24 h of culture. conclusion: Ceftazidime, but not gentamicin or meropenem reduced ureteric branching in mice and suggest a role for Fgf8 and Gdnf in its mechanism. Metanephros surface area measurements can be used to reduce intra-and inter-litter variation. k idney development, leading to the formation of nephrons, starts around the 5th wk of gestation and terminates before term birth, around the 34th-36th wk of gestation. Many factors have been described to disturb this developmental process, leading to long-term problems such as hypertension and chronic kidney disease (1).One such disturbing factor may be the use of (nephrotoxic) drugs during kidney development, such as in pregnant women or neonates born before termination of nephrogenesis. Gentamicin, as well as other aminoglycosides, is widely used as part of the first line treatment of (suspected) bacterial infection in neonates to combat Gram-negative infections. Based on data from the Netherlands Perinatal Registry, 62% of neonates born before 32 wk, who can be considered the most vulnerable group, are treated with aminoglycosides (2). However, due to the fact that gentamicin is classified as a nephrotoxic drug, controversy remains on its safety as aminoglycosides have been shown to disturb kidney development and lead to a reduced nephron number in some experimental animals (3-6) and organ culture studies (7).The aim of our research was to study the impact of antibiotic treatments on nephrogenesis in a model of early nephrogenesis. Gentamicin was studied as well as clinically relevant, alternative drug treatments to compare the toxic potentials of these drugs in a clinical dose range. As alternative treatments, we chose a third generation cephalosporin, ceftazidime, and the carbapenem meropenem. These two drugs both have properties to deal with Gram-negative bacteria and have different mechanisms of action compared to gentamicin. Although beta-lactams have their own potencies to...

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Section: Discussioncontrasting

confidence: 63%

Section: Metanephric Growthmentioning

confidence: 99%

Antibiotics and renal branching morphogenesis: comparison of toxicities

et al. 2014

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“…Recent years have seen the development of a new culture system that allows intact fetal kidney rudiments to produce distinct cortex and medulla, and loops of Henle, in culture (10). In this system, the engineered organs behave similarly (Fig.…”

Section: Engineering a Kidney De Novomentioning

confidence: 99%

Engineered kidneys: principles, progress, and prospects

Davies

Chang

Lawrence

et al. 2014

Advances in Regenerative Biology

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“…There are alternative culture conditions in which the kidney will grow in a more 3D pattern with less flattening (Giuliani et al 2008;Rosines et al 2010), but these have not been used as extensively. Another recently described culture system, in which kidneys are grown directly on glass, in a low volume of medium, allows more extensive growth than in the standard filter cultures (including development of a distinct cortex and medulla), although the kidneys also flatten in this system (Sebinger et al 2010).…”

Section: Kidney Developmentmentioning

confidence: 99%

Imaging Kidney Development

Costantini¹,

Watanabe²,

Lu³

et al. 2011

Cold Spring Harb Protoc

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INTRODUCTIONDevelopment of the kidney involves interactions between several cell lineages and complex morphogenetic processes, such as branching of the ureteric bud (UB) to form the collecting duct system and condensation and differentiation of the mesenchymal progenitors to form the nephron epithelia. One of the advantages of the mouse kidney as an experimental system is that it can develop in culture, from the stage of initial branching of the UB (E11.5) for up to a week (although it achieves the size and degree of development of only an E13.5–E14.5 kidney in vivo). The availability of fluorescent proteins (FPs) has provided powerful tools for visualizing the morphogenesis of specific renal structures in organ cultures. Two categories of genetically modified mice that express FPs are useful for visualizing different cell lineages and developmental processes in these organ cultures: (1) transgenic mice that express a fluorescent reporter in the pattern of a specific gene; and (2) Cre reporter mice, which turn on an FP in cells with Cre recombinase activity (and their daughter cells), used in conjunction with cell type-specific Cre transgenic mice. Here, we describe some of the currently available Cre and FP transgenic lines that are useful for the study of kidney development.

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A Novel, Low-Volume Method for Organ Culture of Embryonic Kidneys That Allows Development of Cortico-Medullary Anatomical Organization

Cited by 57 publications

References 48 publications

Antibiotics and renal branching morphogenesis: comparison of toxicities

Antibiotics and renal branching morphogenesis: comparison of toxicities

Engineered kidneys: principles, progress, and prospects

Imaging Kidney Development

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