Mononuclear cells from a rare blood donor, after freezing under good manufacturing practice conditions, generate red blood cells that recapitulate the rare blood phenotype

Abstract

Background Cultured red blood cells (cRBCs) from cord blood (CB) have been proposed as transfusion products. Whether buffy-coats discarded from blood donations (AB) may be used to generate cRBCs for transfusion has not been investigated. Study Design and Methods Erythroid progenitor cell content and numbers and blood group antigen profiles of erythroblasts (ERYs) and cRBCs generated in Human Erythroid Massive Amplification (HEMA) culture by CB (n=7) and AB (n=33, three females, three males, one AB with rare blood antigens cryopreserved using CB protocols) were compared. Results Variability was observed both in progenitor cell content (2-fold) and number of ERYs generated (1-log) by CB and AB in HEMA. The average progenitor cell contents of the subset of AB and CB analyzed were similar. AB generated numbers of ERYs 3-times lower (p<0.01) than CB in HEMA containing fetal bovine serum but similar to CB in HEMA containing human proteins. Female AB contained 2-times less (p<0.05) erythroid progenitor cells but generated numbers of ERYs similar to those generated by male AB. Cryopreserved AB with a rare blood group phenotype and shipped to another laboratory generated great numbers of ERYs, 90% of which matured into cRBCs. Blood group antigen expression was consistent with the donor genotype for ERYs generated both by CB and AB but concordant with that of native RBCs only for cells derived from AB. Conclusion Buffy-coats from regular donors, including a donor with rare phenotypes stored under conditions established for CB, are not inferior to CB for the generation of cRBCs.

Preclinical rationale for TGF-β inhibition as a therapeutic target for the treatment of myelofibrosis

“…At time 0, the frequency of CD34 + cells in MNC from AB was similar to that of CB, PV and PMF (Fig. 7A,B), consistent with the knowledge that blood donation induces stem/progenitor cell mobilization [39]. In cultures of AB without SB431542, the frequency of CD34 + cells significantly increased by 1.5–2-fold by 24–48h and became barely detectable by 72h.…”

Large-scalein-vitroproduction of red blood cells from human peripheral blood mononuclear cells

“…We have recently shown that 3ml of blood is sufficient to culture enough cells to reprogram erythroblasts to induced pluripotent stem cells (IPSC) 41 . In addition to direct use, the expanded erythroblast can be frozen and defrosted without loss of expansion potential (data not shown), similar to what we previously reported for starting cultures from frozen PBMCs 42 . This introduces flexibility concerning actual production of products.…”

Large-scale in vitro production of red blood cells from human peripheral blood mononuclear cells

“…41 In addition to direct use, the expanded erythroblast can be frozen and defrosted without loss of expansion potential (data not shown), similar to what we previously reported for starting cultures from frozen PBMC. 42 This introduces flexibility concerning actual production of products (eg, with the generation of IPSC form cryopreserved patient material). [43][44][45] The use of adult PBMC also facilitates the availability of starting material and introduces the possibility to culture autologous cRBC.…”