In vitro clinical-grade generation of red blood cells from human umbilical cord blood CD34+ cells.
BACKGROUND
There is no appropriate alternative source of red blood cells (RBCs) to relieve the worsening shortage of blood available for transfusion. Therefore, in vitro generation of clinically available RBCs from hematopoietic stem cells could be a promising new source to supplement the blood supply. However, there have been few studies about the generation of clinical-grade RBCs by coculture on human mesenchymal stem cells (MSCs) and various cytokine supplements, even though the production of pure RBCs requires coculture on stromal cells and proper cytokine supplements.


METHODS
Umbilical cord blood (CB) CD34+ cells were cultured in serum-free medium supplemented with two cytokine sets of stem cell factor (SCF) plus interleukin-3 (IL-3) plus erythropoietin (EPO) and SCF plus IL-3 plus EPO plus thrombopoietin (TPO) plus Flt-3 for 1 week, followed by coculture upon MSCs derived from bone marrow (BM) or CB for 2 weeks.


RESULTS
Almost pure clinical-grade RBCs could be generated by coculturing with CB-MSCs but not BM-MSCs. Expansion fold and enucleation rate were significantly higher in coculture with CB-MSCs than BM-MSCs. Despite a 2.5-fold expansion of erythroblasts in the presence of TPO and Flt-3 for 8 days, the final RBC count was higher without TPO and Flt-3.


CONCLUSIONS
This study is the first report on generating clinical-grade RBCs by in vitro culture with human MSCs and compared effectiveness of several cytokines for RBC production. This provides a useful basis for future production of clinically available RBCs and a model of erythropoiesis that is analogous to the in vivo system.
Baek
Eun Jung
EJ
The Department of Laboratory Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.
Kim
Han-Soo
HS
Kim
Sinyoung
S
Jin
Honglien
H
Choi
Tae-Yeal
TY
Kim
Hyun Ok
HO
D018952:Antigens, CD34 / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods*; D018920:Coculture Techniques; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D017707:Erythrocyte Transfusion; D004912:Erythrocytes / Q000166:cytology*; D041905:Erythroid Cells / Q000166:cytology; D004921:Erythropoietin / Q000494:pharmacology; D005312:Fetal Blood / Q000166:cytology*; D006801:Humans; D016130:Immunophenotyping; D007377:Interleukin-3 / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D019089:Stem Cell Factor / Q000494:pharmacology; D017154:Stromal Cells / Q000166:cytology; D013926:Thrombopoietin / Q000494:pharmacology; D051941:fms-Like Tyrosine Kinase 3 / Q000494:pharmacology
D018952:Antigens, CD34; D016895:Culture Media, Serum-Free; D007377:Interleukin-3; D019089:Stem Cell Factor; D004921:Erythropoietin; D013926:Thrombopoietin; C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3