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Biochemical characterization of density-separated human erythrocytes
Biochimica Et Biophysica Acta
|January 21, 1976
Summary
This study demonstrates a simple method to separate human erythrocytes by density, revealing age-related biochemical changes. However, ATPase activity varied, cautioning against its use as a sole indicator of red blood cell age.
Area of Science:
- Hematology
- Biochemistry
- Cell Biology
Background:
- Human erythrocyte separation is crucial for biochemical analysis.
- Previous methods had limitations in reproducibility and accuracy.
- Murphy's method offers a simple and reproducible approach.
Purpose of the Study:
- To evaluate Murphy's erythrocyte separation method for biochemical analysis.
- To investigate density-dependent biochemical and hematological changes in erythrocytes.
- To assess the reliability of ATPase activity as an age marker.
Main Methods:
- Phthalate ester density centrifugation of erythrocyte fractions obtained by Murphy's method.
- Analysis of various biochemical and hematological parameters.
- Comparison with existing density separation techniques.
Main Results:
- Erythrocytes separated by density, correlating with age.
- Consistent changes observed in hemoglobin, Na+, K+, and other metabolites with increasing density.
- ATPase activity (ouabain-sensitive and Ca2+-stimulated) showed significant variability, irrespective of cell density.
Conclusions:
- Murphy's method effectively fractionates erythrocytes by density (age) in autologous plasma.
- Biochemical markers reliably indicate erythrocyte age, except for ATPase activity.
- ATPase activity is not a dependable indicator of erythrocyte age due to observed variability.