EBV Immortalization of human B lymphocytes separated from small volumes of cryo-preserved whole blood.
Author(s): Amoli MM, Carthy D, Platt H, Ollier WE
Publication: Int J Epidemiol, 2008, Vol. 37 Suppl 1, Page i41-5
PubMed ID: 18381392 PubMed Review Paper? No
Suggested by: ISBER
Purpose of Paper
The purpose of this paper was to assess the viability, purity, and transformability of B lymphocytes obtained from cryopreserved whole blood held at room temperature for approximately 1 day prior to cryopreservation.
Conclusion of Paper
Thawing of 10 whole blood specimens that had been cryopreserved without a delay at room temperature revealed significant haemolysis. The B lymphocytes obtained by magnetic-activated cell sorting were >95% pure by flow cytometry and 70-75% appeared viable by phase contrast microscopy. All 10 specimens were subsequently Epstein Barr Virus (EBV) immortalized. A delay at room temperature of approximately 1 day prior to cryopreservation of blood specimens resulted in 92.5% EBV transformability. The nontransformable specimens consisted of one specimen with a fungal contamination and two others with bacterial contamination.
Studies
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Study Purpose
The purpose of this study was to assess the viability, purity, and transformability of B lymphocytes obtained from cryopreserved whole blood held at room temperature for approximately 1 day prior to cryopreservation.
Summary of Findings:
Thawing of 10 whole blood specimens that had been cryopreserved without a delay at room temperature revealed significant haemolysis. The B lymphocytes obtained by magnetic-activated cell sorting were >95% pure by flow cytometry and 70-75% appeared viable by phase contrast microscopy. All 10 specimens were subsequently EBV immortalized. A delay at room temperature of approximately 1 day prior to cryopreservation of blood specimens resulted in 92.5% EBV transformability. The nontransformed specimens consisted of one specimen with fungal contamination and two others with bacterial contamination.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Normal
Platform:
Analyte Technology Platform Cell count/volume Flow cytometry Cell count/volume Magnetic-activated cell sorting Morphology Macroscopic observation Morphology Colorimetric assay Morphology Light microscopy Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Time at room temperature 0 d
1 d
Storage Storage temperature -150 degrees C
Biospecimen Acquisition Anticoagulant Acid-citrate-dextrose