Immunomagnetic T Cell Depletion: an Analysis of Variables Affecting Final Cell Yield

Clin Lab. 2016 Jul 1;62(7):1243-1248. doi: 10.7754/Clin.Lab.2015.151026.

Abstract

Background: Only little detailed information has been published by a small number of centers on experiences with the technical aspects of "in vitro" large-scale graft manipulation technologies.

Methods: We report our experiences with the graft engineering procedures performed and the results obtained after T cell depletion. We have analyzed data from 212 procedures (108 CD34+ cell selection and 104 CD3+/CD19+ cell depletion).

Results: We conclude that the final cell products after selection or depletion were completely different with regard to CD34+ cell purity (95.8% vs. 1.52%). The CD34+ cell recovery after CD34+ cell selection is negatively affected when the initial leukocyte and/or CD34+ cell counts exceed the threshold defined by the manufacturer (68.9% vs. 45.2%, p < 0.01). However, the cell count threshold defined for the depletion technique could be exceeded without seriously affecting final results (86.1% vs. 86.4% for those with more or less than 40 x 109 leukocyte before the procedure; p = 0.7). Another important conclusion from this study is that in both CD34+ cell selection and CD3+/CD19+ cell depletion better results were reached after having gained experience by performing the procedures several times. This means that a learning process can be expected when using these in vitro graft manipulation procedures.

Conclusions: It is extremely important to have experienced staff to perform these procedures. The expected results are different with each procedure so the decision on which of these T cell depletion approaches are used should be based on the characteristics of the final product wanted.

MeSH terms

  • Allografts
  • Antigens, CD19
  • Antigens, CD34
  • CD3 Complex
  • Flow Cytometry
  • Hematopoietic Stem Cell Transplantation
  • Humans
  • Immunomagnetic Separation*
  • Learning Curve
  • Lymphocyte Count
  • Lymphocyte Depletion / methods*
  • Platelet Count
  • T-Lymphocytes / cytology*
  • T-Lymphocytes / immunology

Substances

  • Antigens, CD19
  • Antigens, CD34
  • CD3 Complex