COLONY FORMING UNITS AS FUNCTIONAL ASSAY FOR RELEASING UMBILICAL CORD BLOOD PRODUCTS FOR HEMATOPOIETIC STEM CELL TRANSPLANT

ROMINA SAINZ; MATIAS GHIGULY; CAROLINA BENTHAM; ELEONORA PAGANO; AGUSTINA MIGUEL; ROSARIO SILVESTRI; JULIETA ROSELLO; CECILIA GAMBA; SILVINA KUPERMAN; VALERIA ROCA

Buenos Aires

Congreso; Reunión Conjunta de Sociedades de Biociencias; 2017

Sociedad Argentina de Investigación Clínica

Umbilical cord blood (CB) is an alternative source of hematopoietic progenitor cells (HPC) for transplant of a great variety of diseases, such as hematological malignancies, marrow failure, immunodeficiencies, hemoglobinopathies and inherited metabolic diseases. Transplantation with CB allows for faster availability, since CB is cryopreserved in banks around the world. However, according to local regulation and international quality standards, cryopreserved products shall be monitored through a stability program and quality control tests should be performed prior to distribution. The facility should define the critical product characteristics to be tested prior to release: viability of an attached segment, confirmatory HLA typing, among others. More than one test might be needed to evaluate the potency of the product prior to its release for transplantation. Colony forming units (CFU) is an in vitro assay developed to support optimal growth of different types of HPC.Our goal was to study CFU assay as a functional test for releasing a CB unit for transplant.We tested viability of CD45+ cells by flow cytometry (FC) with 7-AAD staining (Mean±SD) in 11 CB units before cryopreservation and a thawed segment of each of them. For segments we also performed CFU assay and viability testing of CD34+ cells (Mean±SD) by FC.Viability of CD45+ cells for non cryopreserved samples was 93 ± 7.5% (n=11), and for thawed samples 65 ± 14.7% (n=10). In the segments the viability of CD34+ cells was 75 ± 15.5 % (n=10). However, for one unit it was not possible to analyze the FC results of the attached segment. Instead, all samples tested (n=11) showed HPC colonies growth in CFU assay.Our results showed that CFU assay could be a valuable test to asses CB potency, and could be defined as a release criterion, especially when results cannot be obtained by FC.