Growing and Genetically Manipulating Human Umbilical Cord Blood-Derived Erythroid Progenitor (HUDEP) Cell Lines

Methods Mol Biol. 2018:1698:275-284. doi: 10.1007/978-1-4939-7428-3_17.

Abstract

The recently established human umbilical cord blood-derived erythroid progenitor (HUDEP) cell lines have equipped red blood cell researchers with valuable in vitro models of erythroid development. Of the three established HUDEP cell lines, HUDEP-2 cells express predominantly adult β-globin and most closely resemble adult erythroid cells. This chapter describes culture protocols for the maintenance and erythroid differentiation of HUDEP-2 cells. Methods to genetically manipulate HUDEP-2 cells using a CRISPR/Cas9 nuclease-based approach are also discussed.

Keywords: CRISPR/Cas9; Erythroid differentiation; Fetal hemoglobin; HUDEP; Hemoglobin switching; Immortalized erythroid cell line.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Biomarkers
  • CRISPR-Cas Systems
  • Cell Culture Techniques*
  • Cell Differentiation
  • Cell Line
  • Cell Self Renewal
  • Erythroid Precursor Cells / cytology*
  • Erythroid Precursor Cells / metabolism*
  • Fetal Blood / cytology*
  • Flow Cytometry
  • Genetic Engineering* / methods
  • Genetic Vectors / genetics
  • Hemoglobins / genetics
  • Histocytochemistry
  • Humans
  • Karyotype
  • Lentivirus / genetics
  • Transduction, Genetic

Substances

  • Biomarkers
  • Hemoglobins