Validation of the Efficacy of a Practical Method for Neutrophils Isolation from Peripheral Blood

Jonathan Degel; Masih Shokrani

doi:10.29074/ascls.23.2.94

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Jonathan Degel, BS
1. St. Louis, MO 63127
Masih Shokrani, PhD, MT(ASCP)⇑
1. Northern Illinois University,, DeKalb, Illinois 60116-2828

Address for Correspondence: Masih Shokrani, PhD MT(ASCP), Assistant Professor, Clinical Laboratory Sciences Program, School of Allied Health and Communicative Disorders, College of Health and Human Sciences, Northern Illinois University, DuSable Hall 165, DeKalb, Illinois 60116-2828, mshokrani{at}niu.edu

Abstract

OBJECTIVE: The objectives of this study were to validate the Polymorphprep™ method for neutrophils isolation for conditions within an identified clinical laboratory setting and to confirm the purity and viability of isolated neutrophils utilizing standard clinical laboratory cell counting, differential, and viability methods.

DESIGN: A validation study, utilizing a quantitative and qualitative design, was performed.

PATIENTS /SAMPLES: 18 EDTA tube blood samples were collected from donors between the ages of 18-60, with no underlying condition. The study was performed in a CLS student laboratory of a midwestern university. The Institutional Review Board approved the study and informed consent was obtained from each subject before blood collection.

INTERVENTIONS: Blood samples were treated using a Polymorphprep™ procedure to isolate a pure population of neutrophils.

MAIN OUTCOME MEASURES: Pre and post cell counts were performed using an automated cell counter. Manual white cells differentials and Trypan blue white blood cells viability analysis was performed.

RESULTS: The mean purity levels were at least 94% following isolation with Polymorphprep™, the mean viability of neutrophils was 99%.

CONCLUSION: The use of Polymorphprep™ technique provides a practical and effective method for the isolation of neutrophils from peripheral blood which can be used for investigative purposes. Validation of the purity of the isolated cell population can be readily confirmed using standard clinical laboratory cell counting and differential analysis.

ABBREVIATIONS: CBC= Complete blood count, EDTA= ethylenediaminetetraacetic acid, WBC= White blood Cells, PMN = Polymorphonuclear neutrophil, Peripheral blood mononuclear cells (PBMCs)