Generation of HLA-Universal iPSC-Derived Megakaryocytes and Platelets for Survival Under Refractoriness Conditions
Autor: | Lena Engels, Christiane Gras, Susanne Aufderbeck, Nico Lachmann, Constanca Figueiredo, Christina Wolf, Kai Schulze, Axel Schambach, Carlos A. Guzmán, Ann-Kathrin Börger, Thomas Moritz, Rainer Blasczyk, Dorothee Eicke, Ulrich Martin, Britta Eiz-Vesper |
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Jazyk: | angličtina |
Rok vydání: | 2016 |
Předmět: |
0301 basic medicine
Cell Human leukocyte antigen 030204 cardiovascular system & hematology Biology Epitope lcsh:Biochemistry 03 medical and health sciences 0302 clinical medicine Immune system Genetics medicine Platelet lcsh:QD415-436 Induced pluripotent stem cell Molecular Biology Genetics (clinical) urogenital system lcsh:RM1-950 Molecular medicine In vitro 030104 developmental biology medicine.anatomical_structure lcsh:Therapeutics. Pharmacology Immunology Molecular Medicine Research Article |
Zdroj: | Molecular Medicine, Vol 22, Iss 1, Pp 274-285 (2016) |
ISSN: | 1528-3658 1076-1551 |
Popis: | 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 after in vivo transfusion in a mouse model indicating that they might be used as an alternative to PLT transfusion. Thus, in vitro produced 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. |
Databáze: | OpenAIRE |
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