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| Item Type: | Article |
|---|---|
| Title: | Deep immune-phenotyping of HLA-homozygous iPS-cardiomyocytes by spectral flow cytometry |
| Creators Name: | Maeding, Nicole, Suresh Kundully, Deepika, Steinhuber, Anna, Kriedemann, Nils, Hernandez-Bautista, Carlos A., Martins, Soraia, Hochmann, Sarah, Wolf, Martin, Mayr, Wolfgang, Jungbauer, Christof, Diecke, Sebastian, Tonn, Torsten, Greber, Boris, Zweigerdt, Robert and Strunk, Dirk |
| 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 thenumberof potential recipients. Here we show deep immune-phenotyping of hiPSC-derived cardiomyocytes(iPS-CM)differentiated from four independentiPSClinesinthree 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 immunephenotyping by spectral flow cytometry at the single-cell level. We found that HLA-homozygousiPSCMsexhibit anoverall stable immune-phenotype across HLAhomozygous 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, PDL2 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 |
| Source: | Frontiers in Immunology |
| ISSN: | 1664-3224 |
| Publisher: | Frontiers Media SA |
| Volume: | 17 |
| Page Range: | 1736994 |
| Date: | 4 February 2026 |
| Official Publication: | https://doi.org/10.3389/fimmu.2026.1736994 |
| PubMed: | View item in PubMed |
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