11/25/2023 0 Comments Any human heart title sequenceMore efforts have been taken to optimize the dissociation protocols and single-cell platforms to sequence adult CMs at a single cellular level recently. In earlier studies of heart-related scRNA-seq, experts focused on the embryonic hearts, in which the CMs were accessible for isolation, and their size was small for single-cell selection. However, due to the difficulties in the dissociation of live cardiomyocytes (CMs) and their large and irregular shape, the applications of scRNA-seq in heart tissue have been delayed compared with that in other organs. Moreover, since they are suitable to detect cell subpopulation, droplet-based scRNA-seq platforms are becoming more popular to profile the organ atlas and characterize adult heart non-myocyte populations in recent studies. Alternatively, Drop-seq, InDrop, 10X Genomics Chromium are economical since they have high throughput while the lowest experimental cost per cell, despite the lower sensitivity. ![]() In addition, it is reported that the combination of an optimized digestion protocol and iCell8 Single-Cell system had a relatively high throughput (>20,000 cells), according to a subsequent study. It has been proven that these platforms exhibited higher sensitivity and lower dropout frequency. However, as SMART-seq2 restricts massively parallel sequencing, the reaction chamber-based technologies such as Fludigm C1, iCell8 Single-Cell system show their priority. For example, through a downstream single-cell cDNA library constructed by manual SMART-seq2 in 396 cells, Nomura, et al., has identified different transcriptomic features and related regulators or signaling pathways that encode morphological and functional phenotype between normal and failing hearts. Due to its high efficiency, it is appropriate to be used on discovering isoform or allelic expression. ![]() Specifically, in terms of the major used platforms, Smart-seq2 could detect more genes per cell. and each technique has its own advantages. Main techniques that have been developed worldwide include Fludigm C1, iCell8 Single-Cell system, Illumina Bio-Rad Single-Cell Sequencing Solution, Drop-seq, InDrop, 10X Genomics Chromium system, Microwell-seq, and SMART-seq2, etc. Recent studies have shown that the single-cell sequencing techniques and platforms have been optimized significantly, which allows it to be used on multiple organs and tissues. We also highlighted the potential significance for a better understanding of heart biology and pathology, as well as looked into the prospect of scRNA-seq in further cardiovascular researches.Ģ. Platforms and advanced proto-cols of SCRNA-seq in heart Herein, we summarized the advances of scRNA-seq platforms and underlined the latest scRNA-seq studies in the area of cardiovascular research. In both animal-based researches and human histological studies, scRNA-seq has been revolutionizing the definition of cardiogenesis, normal heart composition, stem-cell differentiation, vasculature development, cardiovascular diseases, and associated immune landscape. Apart from promoting the discovery of novel cell types, this technique enables the interpretation of endogenous principles in cellular gene patterns, the detections of potential cell interactions and the trajectories of cell fate. Recently, the flourishing scRNA-seq has also shed light on the area of cardiovascular research. Moreover, scRNA-seq enables us to profile the whole transcriptomic landscape of various cell groups, identify previously unknown cell subtypes, elucidate the phenotypic transitions, unveil different cell fates, as well as confirm or expand previous knowledge of various organs and different systems. Since single-cell RNA sequencing (scRNA-seq) technique has been applied to several organs/systems, weĬan conduct the research through individual cell-based resolution, decipher integrated cell-map for organs to gain insights into understanding the cellular heterogeneity of diseases and organism biology. Even though diagnosis and therapy of these diseases have been significantly improved along with the cumulative anatomical, cellular and genomic knowledge and techniques, limitations of understanding endogenous cellular heterogeneity and gene interplay within the development of normal hearts and relevant diseases hinder us in deciphering pathogenesis and introducing better therapeutic strategies. In the past few decades, we have been fighting against various common cardiovascular diseases such as coronary heart disease, arrhythmia, cardiomyopathies, heart failure, aortic dissection, and valve diseases. Because of the essential role of the cardiovascular system, any dysfunctions of heart will lead to damaging outcomes. ![]() The mammalian heart is a complex organ, in which the myocardium, conduction system, conduits, and valves are collaborating to maintain systemic circulation.
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