International Journal of Molecular Sciences (Oct 2020)

Genomic Analysis of Hematopoietic Stem Cell at the Single-Cell Level: Optimization of Cell Fixation and Whole Genome Amplification (WGA) Protocol

  • Chiara Carretta,
  • Selene Mallia,
  • Elena Genovese,
  • Sandra Parenti,
  • Sebastiano Rontauroli,
  • Elisa Bianchi,
  • Sebastian Fantini,
  • Stefano Sartini,
  • Lara Tavernari,
  • Enrico Tagliafico,
  • Rossella Manfredini

DOI
https://doi.org/10.3390/ijms21197366
Journal volume & issue
Vol. 21, no. 19
p. 7366

Abstract

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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.

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