GENERATION OF HUMAN ERYTHROID CELLS IN A THREE-DIMENSIONAL MODEL DERIVED FROM HUMAN INDUCED PLURIPOTENT STEM CELLS

@article{aleixo2026generation,
  title = {{GENERATION OF HUMAN ERYTHROID CELLS IN A THREE-DIMENSIONAL MODEL DERIVED FROM HUMAN INDUCED PLURIPOTENT STEM CELLS}},
  author = {Letícia Ribeiro Carvalho ALEIXO and Jacqueline Manoella BARBALHO},
  journal = {Hematology, Transfusion and Cell Therapy},
  year = {2026},
  month = oct,
  doi = {10.1016/j.htct.2026.108112},
  url = {https://doi.org/10.1016/j.htct.2026.108112},
  note = {CC BY},
}

TY  - JOUR
TI  - GENERATION OF HUMAN ERYTHROID CELLS IN A THREE-DIMENSIONAL MODEL DERIVED FROM HUMAN INDUCED PLURIPOTENT STEM CELLS
AU  - Letícia Ribeiro Carvalho ALEIXO
AU  - Jacqueline Manoella BARBALHO
PY  - 2026
DA  - 2026/10/23
JO  - Hematology, Transfusion and Cell Therapy
DO  - 10.1016/j.htct.2026.108112
UR  - https://doi.org/10.1016/j.htct.2026.108112
AB  - Human induced pluripotent stem cells (hiPSCs) have emerged as a promising source for the in vitro generation of erythroid cells, offering an alternative to donor-dependent blood products while overcoming limitations associated with donor availability, storage constraints and sample variability. In addition to their potential applications in transfusion medicine, hiPSC-derived erythroid cells provide relevant models for investigating erythropoiesis, hematological disorders, host-pathogen interactions and drug screening. Despite recent advances, reproducible three-dimensional differentiation systems capable of generating biologically relevant erythroid cells remain limited. This study aimed to develop and characterize a three-dimensional differentiation model for the generation of human erythroid cells from hiPSCs. Human hiPSC lines established in our laboratory (Ethics Approval: CAAE 63167722.0.0000.5272) were aggregated into three-dimensional embryoid bodies (EBs), multicellular aggregates that mimic early embryonic development by recapitulating the formation of the three germ layers and providing a physiologically relevant microenvironment for lineage specification. EBs were differentiated using a sequential serum-free five-stage protocol comprising mesoderm induction with BMP4, VEGF, bFGF and IWP2; hematopoietic commitment with BMP4, VEGF, bFGF, SCF, IGF-2, IBMX and SR1; followed by erythroid induction with SCF, IL-3 and erythropoietin, erythroid expansion and terminal maturation. Cell differentiation was assessed by flow cytometric analysis of CD34, CD43, CD71 and CD235a expression, Romanowsky staining, immunofluorescence for membrane remodeling and enucleation, and functional susceptibility to Plasmodium vivax invasion. The proposed model reproducibly generated erythroid cells displaying progressive maturation. Flow cytometry demonstrated the expected erythroid immunophenotypic progression, characterized by early CD43 expression, progressive acquisition of CD235a and modulation of CD71, reaching approximately 94% CD235a-positive cells at the final stage. Morphological analyses identified proerythroblasts, polychromatic erythroblasts, orthochromatic erythroblasts and enucleated reticulocytes. Immunofluorescence demonstrated membrane remodeling, nuclear polarization and nuclear extrusion, confirming hallmark events of terminal erythropoiesis. Preliminary invasion assays indicated that hiPSC-derived erythroid cells are susceptible to Plasmodium vivax invasion, providing initial evidence of the biological relevance of the proposed model for host-pathogen interaction studies. Together, the immunophenotypic, morphological and functional findings indicate that this three-dimensional differentiation strategy reproduces key features of human erythroid maturation and represents a promising approach for generating biologically relevant erythroid cells in vitro. These results highlight the potential of this hiPSC-derived three-dimensional erythropoiesis platform for investigating erythropoiesis, modeling hematological diseases and studying host-pathogen interactions. Further optimization and molecular characterization will contribute to consolidating its robustness and expanding its translational applications in hematology.
SN  - 2531-1379
SN  - 2531-1387
ER  -