Detection of Residual Donor Erythroid Progenitor Cells after Hematopoietic Stem Cell Transplantation for Patients with Hemoglobinopathies

J Vis Exp. 2017 Sep 6:(127):56002. doi: 10.3791/56002.

Abstract

The presence of incomplete chimerism is noted in a large proportion of patients following bone marrow transplant for thalassemia major or sickle cell disease. This observation has tremendous implications, as subsequent therapeutic immunomodulation strategies can improve clinical outcome. Conventionally, polymerase chain reaction-based analysis of short tandem repeats is used to identify chimerism in donor-derived blood cells. However, this method is restricted to nucleated cells and cannot distinguish between dissociated single-cell lineages. We applied the analysis of short tandem repeats to flow cytometric-sorted hematopoietic progenitor cells and compared this with the analysis of short tandem repeats obtained from selected burst-forming unit - erythroid colonies, both collected from the bone marrow. With this method we are able to demonstrate the different proliferation and differentiation of donor cells in the erythroid compartment. This technique is eligible to complete current monitoring of chimerism in the stem cell transplant setting and thus may be applied in future clinical studies, stem cell research and design of gene therapy trials.

Publication types

  • Video-Audio Media

MeSH terms

  • Erythroid Precursor Cells / cytology*
  • Erythroid Precursor Cells / transplantation*
  • Hematopoietic Stem Cell Transplantation / methods*
  • Hemoglobinopathies / pathology
  • Hemoglobinopathies / therapy*
  • Humans
  • Transplantation Chimera / physiology*
  • Transplantation Conditioning / methods