Standardized human granulocyte isolation using Mono-Poly™ resolving medium: protocol development, validation, and comparison with Lymphoprep™
- 1Faculty of Pharmacy and Pharmaceutical Sciences, College of Health Sciences, University of Alberta, Edmonton, Canada
- 2Department of Drug Technology, Faculty of Medical Technology, Derna, Libya
- 3Li Ka Shing Institute of Virology, Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Canada
Abstract
High purity neutrophil preparations are required for studying neutrophil biology and function. Although density gradient centrifugation is the most commonly used isolation technique, the choice of gradient medium can significantly affect cell purity and downstream applications. The stepwise method of using Mono-Poly™ Resolving Medium (M-PRM) for human peripheral blood granulocyte isolation is presented here alongside the Lymphoprep™ protocol, as these are the two commercially available density-gradient media selected for comparison in this study. Both protocols were optimized and validated by flow cytometric analysis using sequential FSC/SSC gating, singlet discrimination, and CD15-positive granulocyte phenotype identification. The use of M-PRM resulted in significantly higher average granulocyte purity (95.3%) than Lymphoprep™ (73.9%). This comparative protocol provides a practical guide for selecting an appropriate density gradient medium based on experimental requirements and offers a standardized workflow for obtaining granulocyte-enriched cell preparations suitable for downstream immunological and functional studies.
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BibTeX
@article{eldalal2026standardized,
title = {{Standardized human granulocyte isolation using Mono-Poly™ resolving medium: protocol development, validation, and comparison with Lymphoprep™}},
author = {Othman Eldalal and Steven Lockhart and Huan Liu and John-Paul R. Cooper and Dinesh Babu and Arno G. Siraki},
journal = {MethodsX},
year = {2026},
month = oct,
doi = {10.1016/j.mex.2026.104213},
url = {https://doi.org/10.1016/j.mex.2026.104213},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Standardized human granulocyte isolation using Mono-Poly™ resolving medium: protocol development, validation, and comparison with Lymphoprep™
AU - Othman Eldalal
AU - Steven Lockhart
AU - Huan Liu
AU - John-Paul R. Cooper
AU - Dinesh Babu
AU - Arno G. Siraki
PY - 2026
DA - 2026/10/16
JO - MethodsX
DO - 10.1016/j.mex.2026.104213
UR - https://doi.org/10.1016/j.mex.2026.104213
AB - High purity neutrophil preparations are required for studying neutrophil biology and function. Although density gradient centrifugation is the most commonly used isolation technique, the choice of gradient medium can significantly affect cell purity and downstream applications. The stepwise method of using Mono-Poly™ Resolving Medium (M-PRM) for human peripheral blood granulocyte isolation is presented here alongside the Lymphoprep™ protocol, as these are the two commercially available density-gradient media selected for comparison in this study. Both protocols were optimized and validated by flow cytometric analysis using sequential FSC/SSC gating, singlet discrimination, and CD15-positive granulocyte phenotype identification. The use of M-PRM resulted in significantly higher average granulocyte purity (95.3%) than Lymphoprep™ (73.9%). This comparative protocol provides a practical guide for selecting an appropriate density gradient medium based on experimental requirements and offers a standardized workflow for obtaining granulocyte-enriched cell preparations suitable for downstream immunological and functional studies.
SN - 2215-0161
ER -