Biology:Precision cut kidney slices
Precision-cut kidney slices refer to thin sections of the kidney tissue that are prepared with high precision to study kidney functions, drug metabolism or disease processes. Researchers use these slices to study the impact of substances on renal function.[1][2]
Basic Preparation
Creating Precision-cut Kidney Slices (PCKS) is a meticulous process that involves several essential steps. The use of vibratomes is crucial in ensuring the production of precise and high-quality kidney slices for research purposes.[3]
Use of vibratomes
The basic steps involved in preparing PCKS using Compresstome vibratomes include:[4]
- Tissue Selection
- Start by carefully selecting kidney tissue from the desired species, such as rodents or humans, ensuring the ;tissue is of high quality and health.
- Tissue Embedding
- To facilitate slicing and maintain tissue structure, the kidney tissue is typically embedded in a suitable medium, such as agarose or gelatin, into the specimen holder of the Compresstome vibratome.
- Slicing Process
- The vibratome operates by oscillating a blade vertically at high frequencies while the tissue is submerged in a cutting solution. This mechanical oscillation creates thin and precise slices of kidney tissue. Researchers can adjust cutting parameters, such as slice thickness, to meet specific experimental requirements. Typically, PCKS have thicknesses ranging from 200-500µm.
- Post-processing
- Depending on the research objectives, PCLS may undergo additional steps such as washing, culturing, or treatment with substances of interest, such as drugs or stimuli.
Advantages of PCKS
Precision cut kidney slices offer several benefits. Kidney tissue slices can be produced easily in quick time. Researchers can control the tissue thickness. These slices maintain the organ's structural integrity, providing more accurate insights into physiological and pathological processes. Additionally these slices provide researchers a standardised testing so the variability in test results get reduced.[5]
Disadvantages of PCKS
Producing slices is time-consuming and the process is technically challenging. Tissue can get damaged while slicing. Test results will not be accurate with damaged tissue. Maintaining good health of the sliced tissue is also challenging. The slices may not fully capture the complex three-dimensional interactions within the intact organ, limiting the scope of certain studies.[6]
Experimental applications
Precision cut kidney slices are used to study kidney functions, drug metabolism, kidney diseases and toxicology testing.
Study of kidney functions
PCKS are used to analyse normal kidney functions and kidney's interaction with other organs. Researchers can also investigate how different substances are filtered, reabsorbed, or excreted in the kidney.[7]
Drug Metabolism Studies
With PCKS, how drugs are metabolised in the kidney can be investigated. Researchers can investigate the effects of drugs on various metabolic pathways and understand side effects.[8][9]
Toxicity Assessment
PCKS allow researchers to understand the toxicity of substances on the kidney. This is important for detecting potential side effects of drugs or environmental exposures.[10]
References
- ↑ "Precision Cut Kidney Slices (PCKS) in Chronic Kidney Disease (CKD) to Help Select Better Drug Candidates and Bridge the Translational Gap". https://www.sygnaturediscovery.com/news-and-events/blog/precision-cut-kidney-slices-pcks/#:~:text=Precision%20cut%20kidney%20slices%20(PCKS)%20in%20the%20discovery%20cascade%3A,an%20extended%20period%20of%20time.
- ↑ Poosti, Fariba; Pham, Bao Tung; Oosterhuis, Dorenda; Poelstra, Klaas; van Goor, Harry; Olinga, Peter; Hillebrands, Jan-Luuk (2015-01-01). "Precision-cut kidney slices (PCKS) to study development of renal fibrosis and efficacy of drug targeting ex vivo". Disease Models & Mechanisms 8 (10): 1227–1236. doi:10.1242/dmm.020172. ISSN 1754-8411. PMID 26112172. PMC 4610232. http://dx.doi.org/10.1242/dmm.020172.
- ↑ Siwczak, F.; Hiller, C.; Pfannkuche, H.; Schneider, M. R. (2023). "Culture of vibrating microtome tissue slices as a 3D model in biomedical research". Journal of Biological Engineering 17 (1): 36. doi:10.1186/s13036-023-00357-5. PMID 37264444.
- ↑ "Microtome - an overview | ScienceDirect Topics". https://www.sciencedirect.com/topics/neuroscience/microtome.
- ↑ Jensen, Michael Schou; Merrild, Camilla; Nørregaard, Rikke; Olinga, Peter; Mutsaers, Henricus A. M. (2023). "Standardized Protocol for the Preparation of Precision-Cut Kidney Slices: A Translational Model of Renal Fibrosis". Kidney Research. Methods in Molecular Biology. 2664. pp. 123–134. doi:10.1007/978-1-0716-3179-9_9. ISBN 978-1-0716-3178-2. https://link.springer.com/protocol/10.1007/978-1-0716-3179-9_9.
- ↑ "Tissue Slice - an overview | ScienceDirect Topics". https://www.sciencedirect.com/topics/medicine-and-dentistry/tissue-slice#:~:text=A%20major%20limitation%20of%20tissue,use%20a%20race%20against%20time.
- ↑ Genovese, Federica; Kàrpàti, Zsolt S.; Nielsen, Signe H.; Karsdal, Morten A. (2016). "Precision-Cut Kidney Slices as a Tool to Understand the Dynamics of Extracellular Matrix Remodeling in Renal Fibrosis". Biomarker Insights 11: 77–84. doi:10.4137/BMI.S38439. PMID 27257368.
- ↑ Graaf, Inge AM de; Groothuis, Geny MM; Olinga, Peter (2007). "Precision-cut tissue slices as a tool to predict metabolism of novel drugs". Expert Opinion on Drug Metabolism & Toxicology 3 (6): 879–898. doi:10.1517/17425255.3.6.879. PMID 18028031. https://www.tandfonline.com/doi/abs/10.1517/17425255.3.6.879.
- ↑ De Kanter, R.; Olinga, P.; De Jager, M.H; Merema, M.T; Meijer, D.K.F; Groothius, G.M.M (1999). "Organ Slices as an in Vitro Test System for Drug Metabolism in Human Liver, Lung and Kidney". Toxicology in Vitro 13 (4–5): 737–744. doi:10.1016/S0887-2333(99)00047-8. PMID 20654543. https://www.sciencedirect.com/science/article/abs/pii/S0887233399000478.
- ↑ Parrish, Alan R.; Gandolfi, A.Jay; Brendel, Klaus (1995). "Precision-cut tissue slices: Applications in pharmacology and toxicology". Life Sciences 57 (21): 1887–1901. doi:10.1016/0024-3205(95)02176-J. PMID 7475939. https://www.sciencedirect.com/science/article/abs/pii/002432059502176J.
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