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Pan-myeloid Differentiation of Human Cord Blood Derived CD34+ Hematopoietic Stem and Progenitor Cells
Published on: August 9, 2019
Cellular volume and marker expression in human peripheral blood apheresis stem cells
Siddharth Sharma1, Raquel Cabana, Sherry Shariatmadar
1Department of Pathology, University of Miami Miller School of Medicine, Sylvester Cancer Center, Miami, Florida 33101, USA.
Summary
Coulter volume accurately measures cell size, outperforming light scatter. This study used Coulter volume to analyze normal blood cells and hematopoietic progenitor cells, finding correlations between cell volume and stem cell marker expression.
Area of Science:
- Hematology
- Cell Biology
- Biotechnology
Background:
- Forward angle light scatter is a common method for assessing cell volume.
- Coulter volume offers a more accurate measurement of cell volume compared to light scatter.
Purpose of the Study:
- To determine the mean cell volume and diameter of normal human peripheral blood cells and hematopoietic progenitor cells (HPC-A).
- To investigate the correlation between cell volume and the expression of stem cell markers (CD34, CD90, CD117, CD133) in HPC-A samples.
Main Methods:
- Analysis of Coulter volume and two-color fluorescence of peripheral blood and HPC-A samples.
- Samples were processed using different lysis and sedimentation methods (IMMUNOPrep, FACSLysing, Histopaque-1077).
- Cells were stained with CD45-FITC and PE-labeled CD34, CD90, CD117, and CD133 antibodies.
Main Results:
- Coulter volume measurements revealed distinct mean cell volumes and diameters for lymphocytes, monocytes, and granulocytes across different sample preparation methods.
- HPC-A samples prepared with FACSLysing solution showed larger cell volumes compared to those prepared with Histopaque-1077 sedimentation.
- A correlation was observed between mononuclear cell volume in HPC-A samples and the expression of stem cell markers, with CD90+ cells exhibiting the smallest mean volume.
Conclusions:
- Coulter volume is a superior method for assessing cell volume in hematological samples.
- Cell volume correlates with stem cell marker expression, potentially aiding in the identification of stem cells that may not express conventional markers.
