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Genomic Analysis of Hematopoietic Stem Cell at the Single-Cell Level: Optimization of Cell Fixation and Whole Genome Amplification (WGA) Protocol
Single-cell genomics has become the method of choice for the study of heterogeneous cell populations and represents an elective application in defining the architecture and clonal evolution in hematological neoplasms. Reconstructing the clonal evolution of a neoplastic population therefore represent...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7582552/ https://www.ncbi.nlm.nih.gov/pubmed/33036143 http://dx.doi.org/10.3390/ijms21197366 |
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author | Carretta, Chiara Mallia, Selene Genovese, Elena Parenti, Sandra Rontauroli, Sebastiano Bianchi, Elisa Fantini, Sebastian Sartini, Stefano Tavernari, Lara Tagliafico, Enrico Manfredini, Rossella |
author_facet | Carretta, Chiara Mallia, Selene Genovese, Elena Parenti, Sandra Rontauroli, Sebastiano Bianchi, Elisa Fantini, Sebastian Sartini, Stefano Tavernari, Lara Tagliafico, Enrico Manfredini, Rossella |
author_sort | Carretta, Chiara |
collection | PubMed |
description | Single-cell genomics has become the method of choice for the study of heterogeneous cell populations and represents an elective application in defining the architecture and clonal evolution in hematological neoplasms. Reconstructing the clonal evolution of a neoplastic population therefore represents the main way to understand more deeply the pathogenesis of the neoplasm, but it is also a potential tool to understand the evolution of the tumor population with respect to its response to therapy. Pre-analytical phase for single-cell genomics analysis is crucial to obtain a cell population suitable for single-cell sorting, and whole genome amplification is required to obtain the necessary amount of DNA from a single cell in order to proceed with sequencing. Here, we evaluated the impact of different methods of cellular immunostaining, fixation and whole genome amplification on the efficiency and yield of single-cell sequencing. |
format | Online Article Text |
id | pubmed-7582552 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-75825522020-10-29 Genomic Analysis of Hematopoietic Stem Cell at the Single-Cell Level: Optimization of Cell Fixation and Whole Genome Amplification (WGA) Protocol Carretta, Chiara Mallia, Selene Genovese, Elena Parenti, Sandra Rontauroli, Sebastiano Bianchi, Elisa Fantini, Sebastian Sartini, Stefano Tavernari, Lara Tagliafico, Enrico Manfredini, Rossella Int J Mol Sci Article Single-cell genomics has become the method of choice for the study of heterogeneous cell populations and represents an elective application in defining the architecture and clonal evolution in hematological neoplasms. Reconstructing the clonal evolution of a neoplastic population therefore represents the main way to understand more deeply the pathogenesis of the neoplasm, but it is also a potential tool to understand the evolution of the tumor population with respect to its response to therapy. Pre-analytical phase for single-cell genomics analysis is crucial to obtain a cell population suitable for single-cell sorting, and whole genome amplification is required to obtain the necessary amount of DNA from a single cell in order to proceed with sequencing. Here, we evaluated the impact of different methods of cellular immunostaining, fixation and whole genome amplification on the efficiency and yield of single-cell sequencing. MDPI 2020-10-06 /pmc/articles/PMC7582552/ /pubmed/33036143 http://dx.doi.org/10.3390/ijms21197366 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Carretta, Chiara Mallia, Selene Genovese, Elena Parenti, Sandra Rontauroli, Sebastiano Bianchi, Elisa Fantini, Sebastian Sartini, Stefano Tavernari, Lara Tagliafico, Enrico Manfredini, Rossella Genomic Analysis of Hematopoietic Stem Cell at the Single-Cell Level: Optimization of Cell Fixation and Whole Genome Amplification (WGA) Protocol |
title | Genomic Analysis of Hematopoietic Stem Cell at the Single-Cell Level: Optimization of Cell Fixation and Whole Genome Amplification (WGA) Protocol |
title_full | Genomic Analysis of Hematopoietic Stem Cell at the Single-Cell Level: Optimization of Cell Fixation and Whole Genome Amplification (WGA) Protocol |
title_fullStr | Genomic Analysis of Hematopoietic Stem Cell at the Single-Cell Level: Optimization of Cell Fixation and Whole Genome Amplification (WGA) Protocol |
title_full_unstemmed | Genomic Analysis of Hematopoietic Stem Cell at the Single-Cell Level: Optimization of Cell Fixation and Whole Genome Amplification (WGA) Protocol |
title_short | Genomic Analysis of Hematopoietic Stem Cell at the Single-Cell Level: Optimization of Cell Fixation and Whole Genome Amplification (WGA) Protocol |
title_sort | genomic analysis of hematopoietic stem cell at the single-cell level: optimization of cell fixation and whole genome amplification (wga) protocol |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7582552/ https://www.ncbi.nlm.nih.gov/pubmed/33036143 http://dx.doi.org/10.3390/ijms21197366 |
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