Abstract:The kidney function of patients with chronic kidney disease (CKD) is impaired irreversibly. Organ transplantation is the only treatment to restore kidney function in CKD patients. The assessment of new potential therapeutic procedures relies heavily on experimental animal models, but it is limited by its human predictive capacity. In addition, the frequently used two-dimensional in vitro human renal cell models cannot replicate all the features of the in vivo situation. In this study, we developed a three-dime… Show more
“…However, as the volume increases, it is harder for nutrient solution to penetrate inside, so the cells in the sphere center might die due to lack of nutrition. On the basis of our previous results [39], the relatively ideal 3D organoid is about 200-300 μm at diameter, which corresponds to USC-organoids with the seeding density of 3000-6000 cells/well in this experiment. In Fig.…”
Section: Morphology and Optimization Of Usc-organoids Morphology Obsementioning
confidence: 53%
“…2 a, the size of the organoids grew larger ranging from about 160 μm (1000 cells/well) to almost 400 μm (8000 cells/well) with the increasing initial cell density. HKC organoid morphology displayed the same pattern [ 39 ]. Fig.…”
Section: Resultsmentioning
confidence: 99%
“…The 3D organoids from three groups were treated with three drugs respectively, aspirin, penicillin G, and cisplatin, for nephrotoxicity test. Aspirin, penicillin G, and cisplatin (all from Sigma-Aldrich, St. Louis, MO, USA) were chosen for drug toxicity test because they were proved to be excreted by glomerulus and renal tubules, respectively [39]. The working concentration of these three drugs was all 200 μM, and the drug treatment time was 48 h [39].…”
Section: Drug Toxicity Testmentioning
confidence: 99%
“…Aspirin, penicillin G, and cisplatin (all from Sigma-Aldrich, St. Louis, MO, USA) were chosen for drug toxicity test because they were proved to be excreted by glomerulus and renal tubules, respectively [39]. The working concentration of these three drugs was all 200 μM, and the drug treatment time was 48 h [39]. Organoids were then homogenized and tested by γ-glutamyltransferase (GGT) Activity Colorimetric Assay Kit (Sigma-Aldrich).…”
Background: Organoids play an important role in basic research, drug screening, and regenerative medicine. Here, we aimed to develop a novel kind of three-dimensional (3D) organoids generated from urine-derived stem cells (USCs) and to explore whether kidney-specific extracellular matrix (kECM) could enable such organoids for renal function in vitro. Methods: USCs were isolated from human urine samples and cultured with kECM extraction to generate 3D organoids in vitro. Eight densities from 1000 to 8000 cells per organoids were prepared, and both ATP assay and Live/Dead staining were used to determine the optimal USC density in forming organoids and kECM additive concentration. The morphology and histology of as-made organoids were evaluated by hematoxylin and eosin (H.E.) staining, immunofluorescence staining and whole mount staining. Additionally, RT-qPCR was implemented to detect renal-related gene expression. Drug toxicity test was conducted to evaluate the potential application for drug screening. The renal organoids generated from whole adult kidney cells were used as a positive control in multiple assessments. Results: The optimized cell density to generate ideal USC-derived organoids (USC-organoids) was 5000 cells/well, which was set as applying density in the following experiments. Besides, the optimal concentration of kECM was revealed to be 10%. On this condition, Live/Dead staining showed that USC-organoids were well self-organized without significant cell death. Moreover, H.E. staining showed that compact and viable organoids were generated without obvious necrosis inside organoids, which were very close to renal organoids morphologically. Furthermore, specific proximal tubule marker Aquaporin-1 (AQP1), kidney endocrine product erythropoietin (EPO), kidney glomerular markers Podocin and Synaptopodin were detected positively in USC-organoids with kECM. Nephrotoxicity testing showed that aspirin, penicillin G, and cisplatin could exert drug-induced toxicity on USCorganoids with kECM.
“…However, as the volume increases, it is harder for nutrient solution to penetrate inside, so the cells in the sphere center might die due to lack of nutrition. On the basis of our previous results [39], the relatively ideal 3D organoid is about 200-300 μm at diameter, which corresponds to USC-organoids with the seeding density of 3000-6000 cells/well in this experiment. In Fig.…”
Section: Morphology and Optimization Of Usc-organoids Morphology Obsementioning
confidence: 53%
“…2 a, the size of the organoids grew larger ranging from about 160 μm (1000 cells/well) to almost 400 μm (8000 cells/well) with the increasing initial cell density. HKC organoid morphology displayed the same pattern [ 39 ]. Fig.…”
Section: Resultsmentioning
confidence: 99%
“…The 3D organoids from three groups were treated with three drugs respectively, aspirin, penicillin G, and cisplatin, for nephrotoxicity test. Aspirin, penicillin G, and cisplatin (all from Sigma-Aldrich, St. Louis, MO, USA) were chosen for drug toxicity test because they were proved to be excreted by glomerulus and renal tubules, respectively [39]. The working concentration of these three drugs was all 200 μM, and the drug treatment time was 48 h [39].…”
Section: Drug Toxicity Testmentioning
confidence: 99%
“…Aspirin, penicillin G, and cisplatin (all from Sigma-Aldrich, St. Louis, MO, USA) were chosen for drug toxicity test because they were proved to be excreted by glomerulus and renal tubules, respectively [39]. The working concentration of these three drugs was all 200 μM, and the drug treatment time was 48 h [39]. Organoids were then homogenized and tested by γ-glutamyltransferase (GGT) Activity Colorimetric Assay Kit (Sigma-Aldrich).…”
Background: Organoids play an important role in basic research, drug screening, and regenerative medicine. Here, we aimed to develop a novel kind of three-dimensional (3D) organoids generated from urine-derived stem cells (USCs) and to explore whether kidney-specific extracellular matrix (kECM) could enable such organoids for renal function in vitro. Methods: USCs were isolated from human urine samples and cultured with kECM extraction to generate 3D organoids in vitro. Eight densities from 1000 to 8000 cells per organoids were prepared, and both ATP assay and Live/Dead staining were used to determine the optimal USC density in forming organoids and kECM additive concentration. The morphology and histology of as-made organoids were evaluated by hematoxylin and eosin (H.E.) staining, immunofluorescence staining and whole mount staining. Additionally, RT-qPCR was implemented to detect renal-related gene expression. Drug toxicity test was conducted to evaluate the potential application for drug screening. The renal organoids generated from whole adult kidney cells were used as a positive control in multiple assessments. Results: The optimized cell density to generate ideal USC-derived organoids (USC-organoids) was 5000 cells/well, which was set as applying density in the following experiments. Besides, the optimal concentration of kECM was revealed to be 10%. On this condition, Live/Dead staining showed that USC-organoids were well self-organized without significant cell death. Moreover, H.E. staining showed that compact and viable organoids were generated without obvious necrosis inside organoids, which were very close to renal organoids morphologically. Furthermore, specific proximal tubule marker Aquaporin-1 (AQP1), kidney endocrine product erythropoietin (EPO), kidney glomerular markers Podocin and Synaptopodin were detected positively in USC-organoids with kECM. Nephrotoxicity testing showed that aspirin, penicillin G, and cisplatin could exert drug-induced toxicity on USCorganoids with kECM.
Advances in stem cell cultures and human-induced pluripotent stem cells have inculcated interests in a rapidly evolving concept – ”organoids.” These are three-dimensional (3D) structures mimicking some of the phenomena of the real organs at anatomical, multicellular, and functional levels
in vitro
. Organoids have been proven to be better than two-dimensional cell culture in replicating the functionality, architectural, and geometrical features of tissues
in vivo
. Recent advancements have led to the generation of models for organ development and disease, finding applications in the drug discovery, screening of novel compounds, and personalized medicine. Since organoids follow the same natural pathway as the normal tissue or pathology, they can be used to study the expression of various genotypes and phenotypic variations across different species. In the light of these advancements, organoids are now being merged with bioengineering to come up with even better and reliable models to predict the disease progression and effectiveness of precision medicines, few of its important applications. This article discusses the various aspects of this emerging concept along with its uses, both in the present times and near future, with a special focus on pharmacological applications.
Organoids are miniature, highly accurate representations of organs that capture the structure and unique functions of specific organs. Although the field of organoids has experienced exponential growth, driven by advances in artificial intelligence, gene editing, and bioinstrumentation, a comprehensive and accurate overview of organoid applications remains necessary. This review offers a detailed exploration of the historical origins and characteristics of various organoid types, their applications—including disease modeling, drug toxicity and efficacy assessments, precision medicine, and regenerative medicine—as well as the current challenges and future directions of organoid research. Organoids have proven instrumental in elucidating genetic cell fate in hereditary diseases, infectious diseases, metabolic disorders, and malignancies, as well as in the study of processes such as embryonic development, molecular mechanisms, and host–microbe interactions. Furthermore, the integration of organoid technology with artificial intelligence and microfluidics has significantly advanced large‐scale, rapid, and cost‐effective drug toxicity and efficacy assessments, thereby propelling progress in precision medicine. Finally, with the advent of high‐performance materials, three‐dimensional printing technology, and gene editing, organoids are also gaining prominence in the field of regenerative medicine. Our insights and predictions aim to provide valuable guidance to current researchers and to support the continued advancement of this rapidly developing field.
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