Methods of freezing cord blood hematopoietic stem cells.
Author(s): Antoniewicz-Papis J, Lachert E, Janik K, Letowska M, Wozniak J
Publication: Transfusion, 2014, Vol. 54(1), Page 194-202
PubMed ID: 23621822 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine the effects of cryopreservation and hematopoietic stem cell (HSC) isolation method on cell counts and viability of cord blood (CB).
Conclusion of Paper
Only non-significant differences in post-thaw cell counts and recoveries were observed between the CB frozen by placement in a -80 degrees C freezer and the specimens frozen at a controlled rate. White blood cell (WBC) counts and viability were highest in HSC sedimented with 6% hydroxy ethyl starch (HES), and mononuclear cell (MNC) viability was highest in HSC sedimented with 3% gelatin, but this was not significantly higher than those isolated with other methods. MNC counts and the percentage of CD34 positive cells were not significantly affected by isolation method.
Studies
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Study Purpose
The purpose of this study was to determine the effects of cryopreservation and HSC isolation method on cell counts and viability of CB. CB was collected into citrate-phosphate-dextrose and refrigerated for less than 24 h before preparation. After freezing in a -80 degree C freezer or at 1 degree C/min to -50 degrees C followed by -3 degrees C/min to -90 degrees C, CB was transferred to liquid nitrogen vapor for storage. After storage, specimens were thawed at 37 degrees C.
Summary of Findings:
Only non-significant differences in post-thaw cell counts and recoveries were observed between the CB frozen by placement in a -80 degrees C freezer and the specimens frozen at a controlled rate. WBC counts and viability were highest in HSC sedimented with 6% HES, but WBC counts were only significantly higher in specimens sedimented with 6% HES than those sedimented with 1% plasmasteril (p=0.0015), and WBC viability was only significantly higher in specimens sedimented with 6% HES than those sedimented with gelatin (p= 0.0075) or isolated with TF-4 filters (p=0.0018). MNC viability was highest in HSC sedimented with 3% gelatin, but this was not significantly higher than those isolated with other methods. However, MNC viability was slightly but significantly higher in buffy coat isolated specimens than in TF-4 filtered specimens (p=0.0060). MNC counts and the percentage of CD34 positive cells were not significantly affected by isolation method.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Cell count/volume Flow cytometry Cell count/volume Hematology/ auto analyzer Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Cooling or freezing method/ rate At -80 degrees C
1 degree C/min
Biospecimen Aliquots and Components Blood processing method Sedimentation with 1% plasmasteril
Sedimentation with 6% HES
Sedimentation with 3% gelatin
Isolation with TF-4 filters
Buffy coat isolation