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A deep exploration into terminally differentiated cells is required to understand the impairments for RBC generation and the underlying mechanisms. Here, we mapped an atlas of terminally differentiated cells from umbilical cord blood mononuclear cells (UCBMN) and pluripotent stem cells (PSC) and observed their dynamic regulation of erythropoiesis at single\u2010cell resolution. Interestingly, we detected a few progenitor cells and non\u2010erythroid cells from both origins. In PSC\u2010derived erythropoiesis (PSCE), the expression of haemoglobin switch regulators (BCL11A and ZBTB7A) were significantly absent, which could be the restraint for its adult globin expression. We also found that PSCE were less active in stress erythropoiesis than in UCBMN\u2010derived erythropoiesis (UCBE), and explored an agonist of stress erythropoiesis gene, TRIB3, could enhance the expression of adult globin in PSCE. Compared with UCBE, there was a lower expression of epigenetic\u2010related proteins (e.g., CASPASE 3 and UBE2O) and transcription factors (e.g., FOXO3 and TAL1) in PSCE, which might restrict PSCE's enucleation. Moreover, we characterized a subpopulation with high proliferation capacity marked by CD99<jats:sup>high<\/jats:sup> in colony\u2010forming unit\u2010erythroid cells. Inhibition of CD99 reduced the proliferation of PSC\u2010derived cells and facilitated erythroid maturation. Furthermore, CD99\u2013CD99 mediated the interaction between macrophages and erythroid cells, illustrating a mechanism by which macrophages participate in erythropoiesis. This study provided a reference for improving ex vivo RBC generation.<\/jats:p>","DOI":"10.1111\/cpr.13614","type":"journal-article","created":{"date-parts":[[2024,3,19]],"date-time":"2024-03-19T02:05:33Z","timestamp":1710813933000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Decoding human in vitro terminal erythropoiesis originating from umbilical cord blood mononuclear cells and pluripotent stem cells"],"prefix":"10.1111","volume":"57","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8571-2116","authenticated-orcid":false,"given":"Xiaoling","family":"Wang","sequence":"first","affiliation":[{"name":"Stem Cell and Regenerative Medicine Lab Beijing Institute of Radiation Medicine  Beijing PR China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei","family":"Zhang","sequence":"additional","affiliation":[{"name":"Beijing Institute of Genomics &amp; China National Center for Bioinformation 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