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Related Experiment Videos

Proficiency testing experience for viable CD34+ stem cell analysis.

María C López1, David A Lawrence

  • 1The Wadsworth Center, New York State Department of Health, Albany, NY 12201-0509, USA.

Transfusion
|February 27, 2008
PubMed
Summary
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Accurate quantification of viable CD34+ stem cells is crucial for hematopoietic engraftment. Proficiency testing revealed ToPro-3 may overestimate viable cells compared to 7-AAD, necessitating further research for optimal stem cell quantification.

Area of Science:

  • Hematology
  • Stem Cell Biology
  • Clinical Laboratory Science

Background:

  • Successful hematopoietic engraftment relies on precise enumeration of viable CD34+ stem cells.
  • Accurate quantification methods for viable CD34+ stem cells are essential in clinical practice.

Purpose of the Study:

  • To evaluate clinical laboratory performance in quantifying viable CD34+ stem cells through proficiency testing.
  • To compare the efficacy of different viability dyes, specifically 7-aminoactinomycin D (7-AAD) and ToPro-3, in stem cell analysis.

Main Methods:

  • Proficiency testing was conducted by the New York State Department of Health for viable CD34+ stem cells.
  • Participating laboratories analyzed spiked blood samples, with comparative viability assessments using 7-AAD and ToPro-3 dyes.

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Main Results:

  • Laboratories adapted protocols to incorporate viability dyes for CD34+ stem cell quantification.
  • 7-AAD was the predominant viability dye, while ToPro-3 yielded lower viable CD34+ cell counts compared to 7-AAD.

Conclusions:

  • ToPro-3 may stain more cells, potentially including compromised or apoptotic cells, compared to 7-AAD.
  • Further investigation is required to determine if novel dyes like ToPro-3 offer advantages in quantifying viable CD34+ stem cells for engraftment by including apoptotic cells.