Generation of HLA-Universal iPSC-Derived Megakaryocytes and Platelets for Survival Under Refractoriness Conditions.
Platelet (PLT) transfusion is indispensable to maintain homeostasis in thrombocytopenic patients. However, PLT transfusion refractoriness is a common life-threatening condition observed in multitransfused patients. The most frequent immune cause for PLT transfusion refractoriness is the presence of alloantibodies specific for human leukocyte antigen (HLA) class I epitopes. Here, we have silenced the expression of HLA class I to generate a stable HLA-universal induced pluripotent stem cell (iPSC) line that can be used as a renewable cell source for the generation of low immunogenic cell products. The expression of HLA class I was silenced by up to 82% and remained stable during iPSC cultivation. In this study, we have focused on the generation of megakaryocytes (MK) and PLTs from a HLA-universal iPSC source under feeder- and xeno-free conditions. On d 19, differentiation rates of MKs and PLTs with means of 58% and 76% were observed, respectively. HLA-universal iPSC-derived MKs showed polyploidy with DNA contents higher than 4n and formed proPLTs. Importantly, differentiated MKs remained silenced for HLA class I expression. HLA-universal MKs produced functional PLTs. Notably, iPSC-derived HLA-universal MKs were capable to escape antibody-mediated complement- and cellular-dependent cytotoxicity. Furthermore, HLA-universal MKs were able to produce PLTs afterin vivotransfusion in a mouse model indicating that they might be used as an alternative to PLT transfusion. Thus,in vitroproduced low immunogenic MKs and PLTs may become an alternative to PLT donation in PLT-based therapies and an important component in the management of severe alloimmunized patients.
Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.|REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany.
Ann-Kathrin
AK
Börger
Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.|REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany.
Dorothee
D
Eicke
Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.
Christina
C
Wolf
Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.|REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany.
Christiane
C
Gras
Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.
Susanne
S
Aufderbeck
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany.
Kai
K
Schulze
Leibniz Research Laboratories for Biotechnology and Artificial Organs, Hannover, Germany.|REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany.
Lena
L
Engels
Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.
Britta
B
Eiz-Vesper
Institute of Experimental Hematology, Hannover Medical School, Hannover, Germany.|REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany.
Axel
A
Schambach
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany.|REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany.
Carlos A
CA
Guzman
Institute of Experimental Hematology, Hannover Medical School, Hannover, Germany.|REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany.
Nico
N
Lachmann
Institute of Experimental Hematology, Hannover Medical School, Hannover, Germany.|REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany.
Thomas
T
Moritz
Leibniz Research Laboratories for Biotechnology and Artificial Organs, Hannover, Germany.|REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany.
Ulrich
U
Martin
Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.|REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany.
Rainer
R
Blasczyk
Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.|REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany.
Constança
C
Figueiredo