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Isolation Methods of Peripheral Blood Mononuclear Cells in Spanish Biobanks: An Overview.

Author(s): Consuegra I, Rodríguez-Aierbe C, Santiuste I, Bosch A, Martínez-Marín R, Fortuto MA, Díaz T, Martí S, Muñoz-Fernández MÁ

Publication: Biopreserv Biobank, 2017, Vol. 15(4), Page 305-309

PubMed ID: 28398808 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study compared the effects of blood collection tube type and cryopreservation media used on the viability of white cells from peripheral blood processed at seven biobanks of the Spanish Hematic Derivatives Group. Peripheral blood was collected in Vacutainer or LeucoSep tubes by an unspecified method from healthy adult volunteers and patients (adult and pediatric) with inflammatory, metabolic, infectious, and rare diseases. After collection, the biobanks maintained the tubes at room temperature and within 4 h white cells were isolated by density gradient centrifugation.  Following isolation, specimens were cryopreserved with 90% FCS + 10% DMSO at four of seven biobanks, 92.5% FCS + 7.5% DMSO at one biobank, and 50% RPMI + 40% FCS + 10% DMS0 at the remaining two biobanks. White cells were stored in vapor-phase LN for a mean time of 14 months before viability was determined by manual counting on a light microscope or using an automated cell counter.

   Summary of Findings:

   White cell viability was not significantly different between specimens collected in Vacutainer tubes and LeucoSep tubes (74% and 75.5%, respectively, P=0.4768). Post-thaw white cell viabilities for the four biobanks using 90% FCS + 10% DMSO as cryopreservation media were 77%–86% in adult patients. Significantly lower white cell viabilities of 62% and 65%, were found at the two biobanks using 50% RPMI + 40% FCS + 10% DMS0 (P<0.0001). Although slightly lower viability for specimens from adult patients was noted at the biobank that used 92.5% FCS + 7.5% DMSO (74%), viability of white cells from pediatric specimens at that biobank using the same media was higher (84%). The authors conclude that the slightly lower viability in adult patients was not attributable to reducing the percentage DMSO from 10% to 7.5%. White cell viability was significantly higher in healthy adults than those with inflammatory, metabolic, infectious, or rare diseases (P=0.0022).

   Biospecimens

   • Bodily Fluid - Blood

   Preservative Types

   • Frozen
   • Other Preservative

   Diagnoses:

   • Other diagnoses
   • Normal

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Fluorescent microscopy
   Cell count/volume	Light microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Concentration of fixative	90% FCS + 10% DMSO 92.5% FCS + 7.5% DMSO 50% RPMI + 40% FCS + 10% DMS0
   Biospecimen Acquisition	Type of collection container/solution	BD Vacutainer tube LeucoSep tube
   Preaquisition	Patient age	Adult Pediatric
   Preaquisition	Diagnosis/ patient condition	Normal Patients with inflammatory, metabolic, infectious, or rare diseases
   Storage	Short-term storage solution	90% FCS + 10% DMSO 92.5% FCS + 7.5% DMSO 50% RPMI + 40% FCS + 10% DMS0

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