ABSTRACT
Organoid models are used to study kidney physiology, such as the assessment of nephrotoxicity and underlying disease processes. Personalized human pluripotent stem cell-derived kidney organoids are ideal models for compound toxicity studies, but there is a need to accelerate basic and translational research in the field. Here, we developed an automated continuous imaging setup with the "read-on-ski" law of control to maximize temporal resolution with minimum culture plate vibration. High-accuracy performance was achieved: organoid screening and imaging were performed at a spatial resolution of 1.1 μm for the entire multi-well plate under 3 min. We used the in-house developed multi-well spinning device and cisplatin-induced nephrotoxicity model to evaluate the toxicity in kidney organoids using this system. The acquired images were processed via machine learning-based classification and segmentation algorithms, and the toxicity in kidney organoids was determined with 95% accuracy. The results obtained by the automated "read-on-ski" imaging device, combined with label-free and non-invasive algorithms for detection, were verified using conventional biological procedures. Taking advantage of the close-to-in vivo-kidney organoid model, this new development opens the door for further application of scaled-up screening using organoids in basic research and drug discovery.
Subject(s)
Humans , Kidney , Organoids , Pluripotent Stem CellsABSTRACT
Objective To study the effect of recombinant human platelet activating factor acetylhydrolase (rPAF-AH) on candidate plasticity-related gene 15 (CPG15) expression in focal cerebral ischemia rats.Methods Forty-five SD rats were randomly divided into sham operation group,saline group and rPAF-AH group (15 in each group).A middle cerebral artery occlusion model of rats was established for the latter two groups.Each group was divided into 2 d group,7 d group,14 d group.Five rats in each of 2 d group,7 d group,14 d group were used in experiment.The total protein was extracted from coronary sections using homogenate.CPG15 protein expression was detected by Western blot.Results The CPG15 protein expression level was significantly higher in saline group and rPAF-AH group than in sham operation group on days 2,7,14 and reached its peak on day 7,and in rPAF-AH group than in saline group on day 7 (1.48±0.04 vs 1.12±0.05,P＜0.01).Conclusion PAF-AH plays a positive role in neuroplasticity of cerebral ischemia by upregulating the CPG-15 expression in focal cerebral ischemia rats.