Protocol

Stem Cell Heterogeneity

Volume 1516 of the series Methods in Molecular Biology pp 227-241

Date:

In Vitro Culture of Human Hematopoietic Stem Cells in Serum Free Medium and Their Monitoring by Flow Cytometry

  • Marc CloutierAffiliated withRecherche et Développement, Héma-Québec
  • , Christine JobinAffiliated withRecherche et Développement, Héma-QuébecFaculté des sciences et de génie, Biochimie, Microbiologie et Bio-informatique, Université Laval
  • , Carl SimardAffiliated withRecherche et Développement, Héma-Québec
  • , Sonia NéronAffiliated withRecherche et Développement, Héma-QuébecFaculté des sciences et de génie, Biochimie, Microbiologie et Bio-informatique, Université Laval Email author 

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Abstract

Hematopoietic stem cells can be isolated from human blood cells trapped in leukoreduction systems. The leukoreduction systems filters or chambers are usually discarded from routine blood or platelet donations in blood banks around the world. These CD34+ cells are a good source of normal stem cells and can be used as models to characterize the blood stem cells before and after culture in vitro. This chapter contains detailed methodologies for the isolation of stem cells from peripheral blood, the culture of these cells in a medium exempt of animal proteins and for the flow cytometry analysis of the resulting cell population for the characterization of their differentiation

Keywords

Adult CD34+ cells Hematopoietic progenitors Stem cell expansion Animal protein free medium Flow cytometry Multiparametric methods SPADE