MP05-16 THE DEVELOPMENT OF A THREE-DIMENSIONAL KIDNEY SPHEROID MODEL TO STUDY CALCIUM-BASED CRYSTAL FORMATION
Bibliographic record
Abstract
You have accessJournal of UrologyCME1 Apr 2023MP05-16 THE DEVELOPMENT OF A THREE-DIMENSIONAL KIDNEY SPHEROID MODEL TO STUDY CALCIUM-BASED CRYSTAL FORMATION John Chmiel, Wongsakorn Kiattiburut, Aaron Kwong, Priyanka Prabhu, Hassan Razvi, Leonard Luyt, Jeremy Burton, and Jennifer Bjazevic John ChmielJohn Chmiel More articles by this author , Wongsakorn KiattiburutWongsakorn Kiattiburut More articles by this author , Aaron KwongAaron Kwong More articles by this author , Priyanka PrabhuPriyanka Prabhu More articles by this author , Hassan RazviHassan Razvi More articles by this author , Leonard LuytLeonard Luyt More articles by this author , Jeremy BurtonJeremy Burton More articles by this author , and Jennifer BjazevicJennifer Bjazevic More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003216.16AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: An estimated 10% of North Americans suffer from nephrolithiasis. With rising incidence rates, there is a strong need for models to study the pathophysiology of stone disease. In vitro cell culture models are essential for mechanistic and high-throughput analysis of stone formation and drug treatment; however, there are no suitable models that adequately represent the pathophysiology of stone disease. Advancements in cell culture have given rise to three-dimensional cysts, which are hollow spheroid structures that have apical to basal polarization. These culture systems have similar physiochemical environments to in vivo models and can be cultured for the extended times required for stone formation. Here, we developed a novel 3D kidney cyst model that can generate calcium-based kidney stones. METHODS: Madin-Darby canine kidney (MDCK) cells were cultured in suspension in growth factor-reduced Matrigel to generate hollow spheroid cysts. Crystal formation was induced by culturing cysts in a custom crystal-forming media for two weeks. Immunofluorescent staining was used for characterizing and visualizing the three-dimensional structure. Alendronate-fluorescein, a fluorescently labelled bisphosphonate that specifically binds to calcium-containing crystals, was used along with plain polarized light to visualize the crystal structures. RESULTS: Stains for cellular components and structures demonstrated that the luminal spheroids were successfully formed under the culture conditions. The spheroids also have basal-to-apical polarization suggesting that crystals will form on the inside lumen of the hollow cyst. Birefringent structures resembling calcium oxalate dihydrate and calcium phosphate crystals were observed embedded along the apical face of the spheroid and protruding into the lumen. Alendronate-fluorescein staining confirmed the presence of calcium-containing crystals in the lumen of the cyst. CONCLUSIONS: We have demonstrated that hollow MDCK spheroids cultured with crystal-forming media can generate calcium-containing crystals in the spheroid lumen. This model is the first in vitro tissue culture-based model to reliably develop calcium-based crystals and can be used to investigate their pathogenesis or to perform high-throughput drug screening. Furthermore, it paves the way for personalized medicine by building the foundation for a cell system where renal tissue can be harvested and cultured to identify individual intrinsic risk factors that might contribute to stone development. Source of Funding: Canadian Institutes of Health Research (CIHR) CGS-D, The Northeastern Section of the American Urological Association (NSAUA), Lawson Health Research © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e49 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information John Chmiel More articles by this author Wongsakorn Kiattiburut More articles by this author Aaron Kwong More articles by this author Priyanka Prabhu More articles by this author Hassan Razvi More articles by this author Leonard Luyt More articles by this author Jeremy Burton More articles by this author Jennifer Bjazevic More articles by this author Expand All Advertisement PDF downloadLoading ...
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.001 |
| Bibliometrics | 0.001 | 0.000 |
| Science and technology studies | 0.001 | 0.000 |
| Scholarly communication | 0.001 | 0.001 |
| Open science | 0.001 | 0.001 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.008 | 0.002 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".