1. Clonal Analysis of the Neonatal Mouse Heart using Nearest Neighbor Modeling
- Author
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Michael C Thomas, Melanie Bakovic, Diana C Chong, Edwin S. Iversen, Ravi Karra, Devang Thakkar, and Paige DeBenedittis
- Subjects
0301 basic medicine ,General Chemical Engineering ,Computational biology ,030204 cardiovascular system & hematology ,Biology ,Clonal analysis ,Article ,General Biochemistry, Genetics and Molecular Biology ,k-nearest neighbors algorithm ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Lineage tracing ,Rare events ,Animals ,Myocytes, Cardiac ,Cardiomyocyte proliferation ,Cell Proliferation ,General Immunology and Microbiology ,General Neuroscience ,Regeneration (biology) ,Models, Cardiovascular ,Neonatal mouse ,Clone Cells ,030104 developmental biology ,Animals, Newborn ,Developmental biology - Abstract
By replacing lost or dysfunctional myocardium, tissue regeneration is a promising approach to treat heart failure. However, the challenge of detecting bona fide heart regeneration limits the validation of potential regenerative factors. One method to detect new cardiomyocytes is multicolor lineage tracing with clonal analysis. Clonal analysis experiments can be difficult to undertake, because labeling conditions that are too sparse lack sensitivity for rare events such as cardiomyocyte proliferation, and diffuse labeling limits the ability to resolve clones. Presented here is a protocol to undertake clonal analysis of the neonatal mouse heart by using statistical modeling of nearest neighbor distributions to resolve cardiomyocyte clones. This approach enables resolution of clones over a range of labeling conditions and provides a robust analytical approach for quantifying cardiomyocyte proliferation and regeneration. This protocol can be adapted to other tissues and can be broadly used to study tissue regeneration.
- Published
- 2020