Cryopreservation impact on blood progenitor cells: influence of diagnoses, mobilization treatments, and cell concentration.
Author(s): Majado MJ, Salgado-Cecilia G, Blanquer M, Funes C, González-García C, Insausti CL, Parrado A, Morales A, Minguela A, Moraleda JM
Publication: Transfusion, 2011, Vol. 51, Page 799-807
PubMed ID: 20880003 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine the effects of cryopreservation, pre- and post-freezing storage, patient diagnosis and mobilization protocol on the viability and concentration of peripheral blood progenitor cells (PBPCs) obtained by leukapheresis from patients who underwent autologous PBPC transplantation.
Conclusion of Paper
Freezing PBPCs significantly affected viability and numbers of colony-forming unit-granulocyte-macrophages (CFU-GM), granulocytes, monocytes, and lymphocytes, regardless of diagnosis. Freezing PBPCs decreased the number of CD34 positive cells in some groups. Freezing PBPCs in cryotubes rather than cryovials affected viability, the number of CFU-GM, the percentage of CD34 positive cells and the lymphocyte, monocyte and granulocyte counts, but the effects depended on diagnosis and mobilization procedure. Effects of diagnosis and mobilization therapy on PBPC viability, recovery and lymphocyte counts were observed, but neither factor had an effect on the percentage of CD34 positive cells. The authors report that overnight storage of PBPCs on ice prior to cryopreservation rather than immediate cryopreservation and duration of frozen storage had no effects on any of the analytes.
Studies
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Study Purpose
The purpose of this study was to determine the effects of cryopreservation, pre- and post-freezing storage, diagnosis and mobilization protocol on the viability and concentration of PBPCs obtained by leukapheresis from patients who underwent autologous PBPC transplantation. PBPCs were obtained using a continuous blood cell separator and anticoagulated with sodium heparin plus acid citrate dextrose formula A (ACD-A). PBPCs were processed immediately or stored overnight at 4 degrees C before cryopreservation. PBPC were cryopreserved in DMSO, frozen in a controlled rate freezer and stored in liquid nitrogen. PBPCs were thawed in a 40 degrees C water bath, immediately diluted in phosphate buffered saline and placed on ice.
Summary of Findings:
Freezing significantly reduced viability of PBPCs and the number of CFU-GM, regardless of patient diagnosis. When PBPCs from patients with multiple myeloma and patients on the 1.5 g/m2 cyclophosphamide plus 10 mg/kg/day granulocyte¿colony-stimulating factor (Cy+G-CSF) mobilization protocol were frozen in cryotubes rather than bags, viability and the number of CFU-GM were significantly lower, and the lymphocyte counts were significantly higher, but no differences in recovery rates of PBPCs were observed with other patient diagnoses or mobilization procedures. Further, when PBPCs from patients with non-Hodgkin lymphoma, PBPCs from patients on the Cy+G-CSF mobilization protocol, or PBPCs with a higher than the median cell concentration were frozen in cryotubes rather than bags, the recovery of viable cells was lower (p=0.029, p=0.019 and p=0.037, respectively), but no differences in recovery rates were observed with other diagnoses or mobilization procedures. CD34 positive cell counts in PBPCs declined after freezing when the group as a whole was considered (p<0.025), but the difference was only significant in patients on the Cy+G-CSF mobilization protocol (p<0.010). The percentage of CD34 positive cells was lower in PBPCs from patients with multiple myeloma frozen in bags rather than cryotubes (p=0.015). Freezing led to significantly lower granulocyte counts and higher lymphocyte and monocyte counts than in never frozen specimens, and after thawing, specimens frozen in cryotubes had lower monocyte and granulocyte counts than specimens frozen in cryobags (p<0.001, and p=0.017, respectively). Effects of diagnosis and mobilization therapy on viability, recovery and lymphocyte counts were observed, but these factors had no effect on the percentage of CD34 positive cells. The authors report that neither overnight storage of PBPC on ice prior to cryopreservation rather than immediate cryopreservation nor the duration of frozen storage had effects on any of the analytes.
Biospecimens
Preservative Types
- Frozen
- None (Fresh)
Diagnoses:
- Neoplastic - Carcinoma
- Neoplastic - Lymphoma
- Neoplastic - Leukemia
Platform:
Analyte Technology Platform Cell count/volume Hematology/ auto analyzer Cell count/volume Flow cytometry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage duration 0 h
Overnight
1-3 months
Storage Storage temperature On ice
In liquid nitrogen
Storage Type of storage container Cryotube
Cryobag
Preaquisition Diagnosis/ patient condition Non-hodgkin's lymphoma
Hodgkin's disease
Breast cancer
Acute myloid leukemia
Multiple myeloma
Underwent autologous PBPC transplantation
Storage Freeze/thaw cycling 0 cycles
1 cycle
Biospecimen Preservation Type of fixation/preservation Frozen
None (fresh)
Preaquisition Other drugs 10 to 20 mg/kg/day granulocyte-colony-stimulating factor
Chemotherapy and 10 to 20 mg/kg/day granulocyte-colony-stimulating factor
1.5 g/m2 cyclophosphamide and 10 to 20 mg/kg/day granulocyte-colony-stimulating factor