Research & Innovation
Publications
Deep immune-phenotyping of HLA-homozygous iPS-cardiomyocytes by spectral flow cytometry
PMU Authors
Nicole Maeding, Anna Steinhuber, Sarah Hochmann, Martin Wolf, Christof Jungbauer, Dirk Strunk
All Authors
Nicole Maeding, Deepika Suresh Kundully, Anna Steinhuber, Nils Kriedemann, Carlos A. Hernandez-Bautista, Soraia Martins, Sarah Hochmann, Martin Wolf, Wolfgang Mayr, Christof Jungbauer, Sebastian Diecke, Torsten Tonn, Boris Greber, Robert Zweigerdt, Dirk Strunk
Journal association
FRONTIERS IN IMMUNOLOGY
Abstract
Introduction Immunogenicity of allogeneic human induced pluripotent stem cell (hiPSC)-derived transplants limits their applicability in regenerative medicine. Selecting human leukocyte antigen (HLA)-homozygous hiPSC lines could be a mitigation strategy and haplo-matching would profoundly expand the number of potential recipients. Here we show deep immune-phenotyping of hiPSC-derived cardiomyocytes (iPS-CM) differentiated from four independent iPSC lines in three centers under chemically defined conditions.Methods and results Broad immunophenotyping with 354 antibodies revealed differential expression of 101 immune-related molecules between iPS-CM and the parental hiPSC lines. We selected 54 key immune markers for deep immune-phenotyping by spectral flow cytometry at the single-cell level. We found that HLA-homozygous iPSCMs exhibit an overall stable immune-phenotype across HLA-homozygous and heterozygous hiPSC lines indicating a robust differentiation process. HLA-homozygous iPS-CM displayed significantly reduced HLA-ABC levels compared to heterozygous counterparts with an otherwise conserved immune-phenotype. Upon interferon gamma challenge as a surrogate of immune stress responsiveness, iPS-CM significantly upregulated HLA-ABC, -E, -F, PD-L1, PD-L2 and the 'don't eat me' signal CD47. As a proof-of-concept we used this panel to benchmark iPS-CM differentiation across three production sites in this study.Discussion The data indicate generally stable immune-phenotype of iPS-CM produced at three different sites and support feasibility of monitoring iPS-CM identity by spectral flow cytometry.
Keywords
Cardiomyocyte, hiPSC, Immunophenotyping, Regenerative medicine, Spectral flow cytometry, Transplantation immunology