Characteristics of thawed autologous umbilical cord blood.
Author(s): Rosenau EH, Sugrue MW, Haller M, Fisk D, Kelly SS, Chang M, Hou W, Eldjerou L, Slayton W, Cogle CR, Wingard JR
Publication: Transfusion, 2012, Vol. 52, Page 2234-42
PubMed ID: 22321210 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine the effects of blood processing method, frozen storage duration, storage container and volume on total nucleated cell (TNC) recovery from umbilical cord blood (UCB) from infants with type 1 diabetes.
Conclusion of Paper
Small differences in initial and post-thaw TNC recovery were observed between the processing methods, but the study size was not large enough for statistical comparisons between all methods. Specimen volume, storage container and storage duration had no significant effects on TNC recovery. Post-thaw TNC recovery was strongly correlated with the percentage of CD34 positive cells (R=0.70). Frozen storage duration did not impact colony forming unit granulocyte-macrophage (CFU-GM) levels.
Studies
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Study Purpose
The purpose of this study was to determine the effects of blood processing method, frozen storage duration, storage container and volume on TNC recovery from UCB from infants with type 1 diabetes. UCB specimens were left unprocessed or were subjected to red blood cell (RBC) depletion with HES, density gradient separation, or plasma depletion (1 specimen) depending on cord blood bank source. UCB specimens were shipped in liquid nitrogen to the test site and stored in liquid nitrogen vapor until thawing. Specimens were thawed in a 37°C water bath and washed with a dextran human serum albumin (HSA) solution.
Summary of Findings:
The TNC recovery, prior to frozen storage, was highest in the single specimen subjected to plasma depletion (100%), followed by the two unprocessed specimens (95.5%), and the three specimens subjected to RBC depletion (82.0%). TNC recovery was lowest in the seven specimens subjected to density gradient separation (57.7%). Post-thaw TNC recovery was highest in the two unprocessed specimens (94%), followed by the RBC-depleted specimens (87.7%), those subjected to density gradient separation (76.5%), and lastly, the plasma depleted specimen (75%). Post-thaw TNC recovery was strongly correlated with the percentage of CD34 positive cells (R=0.70). The post-thaw percentage of CD34 positive cells were 43.6%, 34.37%, 91.97% and 30.51% for the three RBC depleted, two density gradient processed, two unprocessed, and one plasma depleted specimen, respectively. While TNC recovery increased slightly when specimens were either frozen in cryobags rather than cryovials (77% versus 86%), stored as aliquots of less than 25 mL rather than greater than 25 mL (77% versus 89%), or were stored less than 5 years rather than longer than 5 years (79% versus 84%), these differences were not significant. Frozen storage duration did not impact CFU-GM.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Diabetes Type 1
Platform:
Analyte Technology Platform Cell count/volume Flow cytometry Cell count/volume Light microscopy Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage duration < 5 years
> 5 years
Biospecimen Aliquots and Components Aliquot size/volume < 25 mL
> 25 mL
Biospecimen Aliquots and Components Blood processing method RBC depletion with HES
Density gradient separation
Not processed
Plasma depletion
Storage Freeze/thaw cycling 0 cycles
1 cycle
Storage Type of storage container Cryobag
Cryovial