{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,1]],"date-time":"2025-11-01T13:52:09Z","timestamp":1762005129591,"version":"build-2065373602"},"reference-count":40,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2019,10,19]],"date-time":"2019-10-19T00:00:00Z","timestamp":1571443200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH\/NHLBI","award":["R01HL130764"],"award-info":[{"award-number":["R01HL130764"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>As cell culture methods and stem cell biology have progressed, the in vitro production of cultured RBCs (cRBCs) has emerged as a viable option to produce cells for transfusion or to carry therapeutic cargoes. RBCs produced in culture can be quality-tested either by xeno-transfusion of human cells into immuno-deficient animals, or by transfusion of autologous cells in immuno-competent models. Although murine xeno-transfusion methods have improved, they must be complemented by studies in immuno-competent models. Non-human primates (NHPs) are important pre-clinical, large animal models due to their high biological and developmental similarities with humans, including their comparable hematopoietic and immune systems. Among NHPs, baboons are particularly attractive to validate cRBCs because of the wealth of data available on the characteristics of RBCs in this species that have been generated by past blood transfusion studies. We report here that we have developed a method to produce enucleated cRBCs by differentiation of baboon induced pluripotent stem cells (iPSCs). This method will enable the use of baboons to evaluate therapeutic cRBCs and generate essential pre-clinical data in an immuno-competent, large animal model. Production of the enucleated baboon cRBCs was achieved by adapting the PSC-RED protocol that we previously developed for human cells. Baboon-PSC-RED is an efficient chemically-defined method to differentiate iPSCs into cRBCs that are about 40% to 50% enucleated. PSC-RED is relatively low cost because it requires no albumin and only small amounts of recombinant transferrin.<\/jats:p>","DOI":"10.3390\/cells8101282","type":"journal-article","created":{"date-parts":[[2019,10,21]],"date-time":"2019-10-21T03:40:29Z","timestamp":1571629229000},"page":"1282","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Differentiation of Baboon (Papio anubis) Induced-Pluripotent Stem Cells into Enucleated Red Blood Cells"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9288-1746","authenticated-orcid":false,"given":"Emmanuel N.","family":"Olivier","sequence":"first","affiliation":[{"name":"Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kai","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joshua","family":"Grossman","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nadim","family":"Mahmud","sequence":"additional","affiliation":[{"name":"Division of Hematology\/Oncology, Department of Medicine, University of Illinois College of Medicine, Chicago, IL 60612, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1084-2135","authenticated-orcid":false,"given":"Eric E.","family":"Bouhassira","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,10,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1016\/j.addr.2016.02.007","article-title":"Red blood cells: Supercarriers for drugs, biologicals, and nanoparticles and inspiration for advanced delivery systems","volume":"106","author":"Villa","year":"2016","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1007\/BF00195932","article-title":"Improved pharmacodynamics of l-asparaginase-loaded in human red blood cells","volume":"49","author":"Kravtzoff","year":"1996","journal-title":"Eur. J. Clin. Pharmacol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.tmrv.2016.08.004","article-title":"Erythrocytes as Carriers for Drug Delivery in Blood Transfusion and Beyond","volume":"31","author":"Villa","year":"2017","journal-title":"Transfus. Med. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1111\/j.1365-2141.2010.08561.x","article-title":"Elimination of blood group antigens: Hope and reality","volume":"152","author":"Bagnis","year":"2011","journal-title":"Br. J. Haematol."},{"key":"ref_5","first-page":"108","article-title":"Use of voltage pulses for the pore opening and drug loading, and the subsequent resealing of red blood cells","volume":"51","author":"Tsong","year":"1985","journal-title":"Bibl. Haematol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.jconrel.2003.11.018","article-title":"Drug, enzyme and peptide delivery using erythrocytes as carriers","volume":"95","author":"Marinero","year":"2004","journal-title":"J. Control. Release"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"891","DOI":"10.1038\/nbt846","article-title":"Prophylactic fibrinolysis through selective dissolution of nascent clots by tPA-carrying erythrocytes","volume":"21","author":"Murciano","year":"2003","journal-title":"Nat. Biotechnol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"4779","DOI":"10.1182\/blood-2011-12-398149","article-title":"Targeting recombinant thrombomodulin fusion protein to red blood cells provides multifaceted thromboprophylaxis","volume":"119","author":"Zaitsev","year":"2012","journal-title":"Blood"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"10131","DOI":"10.1073\/pnas.1409861111","article-title":"Engineered red blood cells as carriers for systemic delivery of a wide array of functional probes","volume":"111","author":"Shi","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1038\/s41467-017-00448-0","article-title":"Genetically engineered red cells expressing single domain camelid antibodies confer long-term protection against botulinum neurotoxin","volume":"8","author":"Huang","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1002\/ajh.24093","article-title":"A Phase 2 study of L-asparaginase encapsulated in erythrocytes in elderly patients with Philadelphia chromosome negative acute lymphoblastic leukemia: The GRASPALL\/GRAALL-SA2-2008 study","volume":"90","author":"Leguay","year":"2015","journal-title":"Am. J. Hematol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.tmrv.2017.09.001","article-title":"A Study of the Pharmacokinetic Properties and the In Vivo Kinetics of Erythrocytes Loaded With Dexamethasone Sodium Phosphate in Healthy Volunteers","volume":"32","author":"Coker","year":"2018","journal-title":"Transfus. Med. Rev."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"10716","DOI":"10.1073\/pnas.191362598","article-title":"Hematopoietic colony-forming cells derived from human embryonic stem cells","volume":"98","author":"Kaufman","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"860","DOI":"10.1182\/blood-2004-11-4522","article-title":"Hematopoietic differentiation of human embryonic stem cells progresses through sequential hematoendothelial, primitive, and definitive stages resembling human yolk sac development","volume":"106","author":"Zambidis","year":"2005","journal-title":"Blood"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Qiu, C., Hanson, E., Olivier, E., Inada, M., Kaufman, D.S., Gupta, S., and Bouhassira, E.E. (2005). Differentiation of human embryonic stem cells into hematopoietic cells by coculture with human fetal liver cells recapitulates the globin switch that occurs early in development. Exp. Hematol., 33.","DOI":"10.1016\/j.exphem.2005.09.003"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1002\/biot.201200368","article-title":"Large-scale production of red blood cells from stem cells: What are the technical challenges ahead?","volume":"9","author":"Rousseau","year":"2014","journal-title":"Biotechnol. J."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1038\/mt.2013.201","article-title":"Extensive ex vivo expansion of functional human erythroid precursors established from umbilical cord blood cells by defined factors","volume":"22","author":"Huang","year":"2014","journal-title":"Mol. Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"604","DOI":"10.5966\/sctm.2012-0059","article-title":"Novel, high-yield red blood cell production methods from CD34-positive cells derived from human embryonic stem, yolk sac, fetal liver, cord blood, and peripheral blood","volume":"1","author":"Olivier","year":"2012","journal-title":"Stem Cells Transl. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2400","DOI":"10.1182\/blood-2007-07-102087","article-title":"Globin switches in yolk sac-like primitive and fetal-like definitive red blood cells produced from human embryonic stem cells","volume":"111","author":"Qiu","year":"2008","journal-title":"Blood"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1541","DOI":"10.1002\/stem.2335","article-title":"\u03b2-Globin-Expressing Definitive Erythroid Progenitor Cells Generated from Embryonic and Induced Pluripotent Stem Cell-Derived Sacs","volume":"34","author":"Fujita","year":"2016","journal-title":"Stem Cells"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.exphem.2019.05.006","article-title":"PSC-RED and MNC-RED: Albumin-free and low-transferrin robust erythroid differentiation protocols to produce human enucleated red blood cells","volume":"75","author":"Olivier","year":"2019","journal-title":"Exp. Hematol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1034","DOI":"10.1016\/j.stemcr.2017.08.018","article-title":"Complement Depletion Improves Human Red Blood Cell Reconstitution in Immunodeficient Mice","volume":"9","author":"Chen","year":"2017","journal-title":"Stem Cell Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/S0070-2153(08)60246-X","article-title":"4 Primate Embryonic Stem Cells","volume":"38","author":"Thomson","year":"1997","journal-title":"Curr. Top. Dev. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1002\/dvdy.1191","article-title":"Establishment of embryonic stem cell lines from cynomolgus monkey blastocysts produced by IVF or ICSI","volume":"222","author":"Suemori","year":"2001","journal-title":"Dev. Dyn."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1530\/REP-09-0067","article-title":"Characterization of a novel embryonic stem cell line from an ICSI-derived blastocyst in the African green monkey","volume":"139","author":"Shimozawa","year":"2010","journal-title":"Reproduction"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"178","DOI":"10.1016\/j.scr.2009.02.004","article-title":"Establishment and characterization of baboon embryonic stem cell lines: An Old World Primate model for regeneration and transplantation research","volume":"2","author":"Simerly","year":"2009","journal-title":"Stem Cell Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1089\/cell.2012.0093","article-title":"Derivation of Induced Pluripotent Stem Cells from the Baboon: A Nonhuman Primate Model for Preclinical Testing of Stem Cell Therapies","volume":"15","author":"Navara","year":"2013","journal-title":"Cell. Reprogram."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1016\/j.stem.2008.10.014","article-title":"Generation of Induced Pluripotent Stem Cells from Adult Rhesus Monkey Fibroblasts","volume":"3","author":"Liu","year":"2008","journal-title":"Cell Stem Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1225","DOI":"10.1089\/scd.2017.0069","article-title":"Induced Pluripotent Stem Cell-Derived Dopaminergic Neurons from Adult Common Marmoset Fibroblasts","volume":"26","author":"Vermilyea","year":"2017","journal-title":"Stem Cells Dev."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1089\/scd.2010.0343","article-title":"Efficient Generation of Nonhuman Primate Induced Pluripotent Stem Cells","volume":"20","author":"Zhong","year":"2011","journal-title":"Stem Cells Dev."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1038\/nmeth.1593","article-title":"Chemically defined conditions for human iPSC derivation and culture","volume":"8","author":"Chen","year":"2011","journal-title":"Nat. Methods"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1111\/j.1939-165X.1999.tb01057.x","article-title":"Rapid staining techniques in cytopathology: A review and comparison of modified protocols for hematoxylin and eosin, Papanicolaou and Romanowsky stains","volume":"28","author":"Lumsden","year":"1999","journal-title":"Vet. Clin. Pathol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1385\/1-59259-373-9:213","article-title":"Transgenic Mice and Hemoglobinopathies","volume":"Volume 82","author":"Fabry","year":"2003","journal-title":"Hemoglobin Disorders"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Navara, C.S., Chaudhari, S., and McCarrey, J.R. (2018). Optimization of culture conditions for the derivation and propagation of baboon (Papio anubis) induced pluripotent stem cells. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0193195"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"877","DOI":"10.1016\/S1095-6433(02)00205-2","article-title":"Globin gene switching in primates","volume":"133","author":"Johnson","year":"2002","journal-title":"Comp. Biochem. Physiol. A Mol. Integr. Physiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1182\/blood.V53.1.19.19","article-title":"Fetal hemoglobin (HbF) synthesis in baboons, Papio cynocephalus. Analysis of fetal and adult hemoglobin synthesis during fetal development","volume":"53","author":"Mueller","year":"1979","journal-title":"Blood"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.celrep.2019.02.021","article-title":"BE Ultra-High Frequency Reprogramming Of Individual Long-Term-Hematopoietic Stem Cells Yields Low Somatic Variant Induced-Pluripotent Stem Cells","volume":"26","author":"Wang","year":"2019","journal-title":"Cell Rep."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"VandeBerg, J.L., Williams-Blangero, S., and Tardif, S.D. (2009). The Baboon in Biomedical Research, Springer.","DOI":"10.1007\/978-0-387-75991-3"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/j.1537-2995.2006.00922.x","article-title":"The survival and function of baboon red blood cells, platelets, and plasma proteins: A review of the experience from 1972 to 2002 at the Naval Blood Research Laboratory, Boston, Massachusetts","volume":"46","author":"Valeri","year":"2006","journal-title":"Transfusion"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Mahmud, N., Khanal, A., Taioli, S., Koca, E., Gaitonde, S., Petro, B., Sweiss, K., Halliday, L., Wang, X., and Patel, P. (2018). Preclinical IV busulfan dose-finding study to induce reversible myeloablation in a non-human primate model. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0206980"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/8\/10\/1282\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:27:49Z","timestamp":1760189269000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/8\/10\/1282"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,10,19]]},"references-count":40,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2019,10]]}},"alternative-id":["cells8101282"],"URL":"https:\/\/doi.org\/10.3390\/cells8101282","relation":{},"ISSN":["2073-4409"],"issn-type":[{"type":"electronic","value":"2073-4409"}],"subject":[],"published":{"date-parts":[[2019,10,19]]}}}