An objective flow cytometry method to rapidly determine cord blood potency in cryopreserved units.
Transfusion
; 59(6): 2074-2083, 2019 06.
Article
en En
| MEDLINE
| ID: mdl-30802338
ABSTRACT
BACKGROUND:
Cord blood banks have to determine the regenerative potential of cord blood units (CBUs) on a representative sample of the cryopreserved product before release to the transplant center. Potency can be measured by using a colony-forming unit (CFU) method, which delays the release of CBU by 7 to 14 days. To accelerate CBU qualification, we have developed a rapid method to assess the response of CD34 cells to interleukin (IL)-3. Flow cytometry was used to measure IL-3-induced STAT5 phosphorylation within CD34-cells. This IL-3 test was compared to the CFU method, as well as the aldehyde dehydrogenase (ALDH) enzyme-based assay. STUDY DESIGN ANDMETHODS:
Ten cryopreserved CBUs were analyzed for their contents in CD34 and CD45 viable cells, total CFUs, ADLHbright cells, and IL-3-responsive CD34+ cells. Extreme and mild warming event scenarios were simulated on CBUs and used as poor-quality samples. Segments, tubes, and bags from five CBUs were compared for their potency using IL-3 and CFU methods.RESULTS:
The IL-3 test was accurate in identifying the samples handled following standard operating procedures and those subjected to extreme warming events. Based on these results, a threshold of 55% of IL-3-responsive CD34 cells was established to identify good-quality samples. The IL-3 test was also the most sensitive to detect samples subjected to milder warming events.CONCLUSIONS:
Our new method for determining CBU functionality is rapid, unbiased, and robust. The IL-3 test described herein fulfills the requirements for validation, and we intend to implement this method in our cord blood bank facility.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Trasplante de Células Madre de Sangre del Cordón Umbilical
/
Sangre Fetal
/
Citometría de Flujo
Tipo de estudio:
Prognostic_studies
Límite:
Female
/
Humans
/
Newborn
/
Pregnancy
Idioma:
En
Revista:
Transfusion
Año:
2019
Tipo del documento:
Article
País de afiliación:
Canadá