{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,15]],"date-time":"2026-06-15T05:05:48Z","timestamp":1781499948679,"version":"3.54.1"},"reference-count":69,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T00:00:00Z","timestamp":1766016000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,12,19]],"date-time":"2025-12-19T00:00:00Z","timestamp":1766102400000},"content-version":"vor","delay-in-days":1,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"Universit\u00e4tsklinikum D\u00fcsseldorf. Anstalt \u00f6ffentlichen Rechts"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Pre-clinical research on B and NK cell development relies on murine stromal cell-based systems with reduced physiological relevance and clinical applicability. A serum-free, fully humanized co-culture system utilizing human bone marrow-derived mesenchymal stromal cells (BM-MSCs) was developed to differentiate CB-CD34+\u2009cells towards B and NK cell lineages. Differentiation dynamics were monitored via flow cytometry, with immunophenotypic analysis tracking progression from progenitors to mature cells. The system generated CD19+\u2009IgM+\u2009immature B cells and CD56+\u2009CD16+\u2009NK cells, recapitulating fetal stages of human lymphopoiesis. Serum-free media conditions ensured reproducibility and high overall yield of CD19+\u2009B (35\u2009\u00b1\u20095.32%) and CD56+\u00a0NK (28.46\u2009\u00b1\u20097.01%) cell progenitors. Flow cytometry identified distinct population peaks, confirming temporal control over differentiation. This clinically relevant platform addresses the limitations of traditional models by providing a more physiologically accurate human microenvironment. The serum-free system supports applications in disease modeling, genotoxic compound screening, and mutational studies of hematopoiesis. By enabling scalable production of B and NK cells it aims to accelerate translational research for immunodeficiencies, cancer immunotherapy, and hematopoietic disorders.<\/jats:p>","DOI":"10.1038\/s41598-025-30732-9","type":"journal-article","created":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T21:45:32Z","timestamp":1766094332000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Serum-free differentiation platform for the generation of B lymphocytes and natural killer cells from human CD34+\u2009cord blood progenitors"],"prefix":"10.1038","volume":"15","author":[{"given":"Rigveda","family":"Bhave","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carla-Johanna","family":"Kath","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nadine","family":"R\u00fcchel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eleni","family":"Vasileiou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vera H.","family":"Jepsen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katharina","family":"Raba","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aleksandra A.","family":"Pandyra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ute","family":"Fischer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gesine","family":"Kogler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,12,18]]},"reference":[{"issue":"2","key":"30732_CR1","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1016\/j.stem.2012.01.006","volume":"10","author":"S Doulatov","year":"2012","unstructured":"Doulatov, S., Notta, F., Laurenti, E. & Dick, J. E. Hematopoiesis: A human perspective. Cell Stem Cell 10(2), 120\u2013136 (2012).","journal-title":"Cell Stem Cell"},{"issue":"6039","key":"30732_CR2","first-page":"218","volume":"333","author":"F Notta","year":"2011","unstructured":"Notta, F. et al. Isolation of single human hematopoietic stem cells capable of long-term multilineage engraftment. Science. (1979) 333(6039), 218\u2013221 (2011).","journal-title":"Science. (1979)"},{"issue":"3","key":"30732_CR3","doi-asserted-by":"publisher","first-page":"326","DOI":"10.15283\/IJSC19157","volume":"13","author":"P Yadav","year":"2020","unstructured":"Yadav, P., Vats, R., Bano, A. & Bhardwaj, R. Hematopoietic stem cells culture, expansion and differentiation: An insight into variable and available media. Int. J. Stem Cells. 13(3), 326. https:\/\/doi.org\/10.15283\/IJSC19157 (2020).","journal-title":"Int. J. Stem Cells."},{"issue":"4","key":"30732_CR4","doi-asserted-by":"publisher","first-page":"491","DOI":"10.1182\/BLOOD-2013-02-453175","volume":"122","author":"KK Ballen","year":"2013","unstructured":"Ballen, K. K., Gluckman, E. & Broxmeyer, H. E. Umbilical cord blood transplantation: The first 25 years and beyond. Blood 122(4), 491\u2013498. https:\/\/doi.org\/10.1182\/BLOOD-2013-02-453175 (2013).","journal-title":"Blood"},{"issue":"6","key":"30732_CR5","doi-asserted-by":"publisher","first-page":"317","DOI":"10.1097\/MOH.0000000000000732","volume":"29","author":"PA Shi","year":"2022","unstructured":"Shi, P. A., Luchsinger, L. L., Greally, J. M. & Delaney, C. S. Umbilical cord blood: An undervalued and underutilized resource in allogeneic hematopoietic Stem cell transplant and novel cell therapy applications. Curr. Opin. Hematol. 29(6), 317. https:\/\/doi.org\/10.1097\/MOH.0000000000000732 (2022).","journal-title":"Curr. Opin. Hematol."},{"issue":"1","key":"30732_CR6","doi-asserted-by":"publisher","first-page":"48","DOI":"10.1038\/s41409-019-0546-9","volume":"55","author":"H Mayani","year":"2019","unstructured":"Mayani, H., Wagner, J. E. & Broxmeyer, H. E. Cord blood research, banking, and transplantation: achievements, challenges, and perspectives. Bone Marrow Transplant 55(1), 48\u201361. https:\/\/doi.org\/10.1038\/s41409-019-0546-9 (2019).","journal-title":"Bone Marrow Transplant"},{"key":"30732_CR7","doi-asserted-by":"publisher","first-page":"1167266","DOI":"10.3389\/FONC.2023.1167266","volume":"13","author":"J Wang","year":"2023","unstructured":"Wang, J. & Metheny, L. Umbilical cord blood derived cellular therapy: advances in clinical development. Front. Oncol. 13, 1167266. https:\/\/doi.org\/10.3389\/FONC.2023.1167266 (2023).","journal-title":"Front. Oncol."},{"issue":"2","key":"30732_CR8","doi-asserted-by":"publisher","first-page":"415","DOI":"10.1038\/leu.2014.215","volume":"29","author":"HJ Pegram","year":"2014","unstructured":"Pegram, H. J. et al. IL-12-secreting CD19-targeted cord blood-derived T cells for the immunotherapy of B-cell acute lymphoblastic leukemia. Leukemia 29(2), 415\u2013422. https:\/\/doi.org\/10.1038\/leu.2014.215 (2014).","journal-title":"Leukemia"},{"key":"30732_CR9","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2024.101361","author":"WL Liu","year":"2024","unstructured":"Liu, W. L. et al. Off-the-shelf allogeneic natural killer cells for the treatment of COVID-19. Mol. Ther. Methods Clin. Dev. https:\/\/doi.org\/10.1016\/j.omtm.2024.101361 (2024).","journal-title":"Mol. Ther. Methods Clin. Dev."},{"issue":"5","key":"30732_CR10","doi-asserted-by":"publisher","first-page":"598","DOI":"10.1016\/j.exphem.2008.01.001","volume":"36","author":"V McCullar","year":"2008","unstructured":"McCullar, V. et al. Mouse fetal and embryonic liver cells differentiate human umbilical cord blood progenitors into CD56-negative natural killer cell precursors in the absence of interleukin-15. Exp. Hematol. 36(5), 598\u2013608. https:\/\/doi.org\/10.1016\/j.exphem.2008.01.001 (2008).","journal-title":"Exp. Hematol."},{"key":"30732_CR11","doi-asserted-by":"publisher","DOI":"10.1016\/J.XPRO.2021.100420","author":"SE Richardson","year":"2021","unstructured":"Richardson, S. E., Ghazanfari, R., Chhetri, J., Enver, T. & B\u00f6iers, C. In vitro differentiation of human pluripotent stem cells into the B lineage using OP9-MS5 co-culture. STAR Protoc. https:\/\/doi.org\/10.1016\/J.XPRO.2021.100420 (2021).","journal-title":"STAR Protoc."},{"issue":"12","key":"30732_CR12","doi-asserted-by":"publisher","first-page":"3683","DOI":"10.1182\/BLOOD.V97.12.3683","volume":"97","author":"QL Hao","year":"2001","unstructured":"Hao, Q. L. et al. Identification of a novel, human multilymphoid progenitor in cord blood. Blood 97(12), 3683\u20133690. https:\/\/doi.org\/10.1182\/BLOOD.V97.12.3683 (2001).","journal-title":"Blood"},{"key":"30732_CR13","doi-asserted-by":"publisher","first-page":"855","DOI":"10.1002\/(SICI)1521-4141(199803)28:03<855::AID-IMMU855>3.0.CO;2-X","volume":"28","author":"M Nishihara","year":"1998","unstructured":"Nishihara, M. et al. A combination of stem cell factor and granulocyte colony-stimulating factor enhances the growth of human progenitor B cells supported by murine stromal cell line MS-5. Eur. J. Immunol. 28, 855\u2013864 (1998).","journal-title":"Eur. J. Immunol."},{"issue":"13","key":"30732_CR14","doi-asserted-by":"publisher","first-page":"1059","DOI":"10.1182\/BLOOD.2019001289","volume":"134","author":"S O\u2019Byrne","year":"2019","unstructured":"O\u2019Byrne, S. et al. Discovery of a CD10-negative B-progenitor in human fetal life identifies unique ontogeny-related developmental programs. Blood 134(13), 1059\u20131071. https:\/\/doi.org\/10.1182\/BLOOD.2019001289 (2019).","journal-title":"Blood"},{"issue":"5","key":"30732_CR15","doi-asserted-by":"publisher","first-page":"587","DOI":"10.1016\/j.exphem.2007.12.013","volume":"36","author":"M Ichii","year":"2008","unstructured":"Ichii, M. et al. Regulation of human B lymphopoiesis by the transforming growth factor-\u03b2 superfamily in a newly established coculture system using human mesenchymal stem cells as a supportive microenvironment. Exp Hematol. 36(5), 587\u2013597. https:\/\/doi.org\/10.1016\/j.exphem.2007.12.013 (2008).","journal-title":"Exp Hematol."},{"issue":"1","key":"30732_CR16","doi-asserted-by":"publisher","first-page":"367","DOI":"10.1182\/BLOOD-2005-07-2657","volume":"107","author":"A Corcione","year":"2006","unstructured":"Corcione, A. et al. Human mesenchymal stem cells modulate B-cell functions. Blood 107(1), 367\u2013372. https:\/\/doi.org\/10.1182\/BLOOD-2005-07-2657 (2006).","journal-title":"Blood"},{"key":"30732_CR17","doi-asserted-by":"publisher","first-page":"47","DOI":"10.3324\/haematol.2009.008524","volume":"95","author":"A Gothot","year":"2010","unstructured":"Gothot, A. et al. Prolonged ex vivo culture of human bone marrow mesenchymal stem cells influences their supportive activity toward NOD\/SCID-repopulating cells and committed progenitor cells of B lymphoid and myeloid lineages. Haematologica 95, 47\u201356. https:\/\/doi.org\/10.3324\/haematol.2009.008524 (2010).","journal-title":"Haematologica"},{"issue":"6","key":"30732_CR18","doi-asserted-by":"publisher","first-page":"20740","DOI":"10.1371\/journal.pone.0020740","volume":"6","author":"J Spanholtz","year":"2011","unstructured":"Spanholtz, J., Preijers, F., Tordoir, M., Trilsbeek, C. & Paardekooper, J. clinical-grade generation of active NK cells from cord blood hematopoietic progenitor cells for immunotherapy using a closed-system culture process. PLoS ONE 6(6), 20740. https:\/\/doi.org\/10.1371\/journal.pone.0020740 (2011).","journal-title":"PLoS ONE"},{"key":"30732_CR19","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2024.101361","author":"WL Liu","year":"2024","unstructured":"Liu, W. L. et al. Off-the-shelf allogeneic natural killer cells for the treatment of COVID-19. Mol. Ther. Methods Clin. Dev. https:\/\/doi.org\/10.1016\/j.omtm.2024.101361 (2024).","journal-title":"Mol. Ther. Methods Clin. Dev."},{"issue":"Supplement 1","key":"30732_CR20","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1182\/BLOOD-2020-142813","volume":"136","author":"D Chandrasekaran","year":"2020","unstructured":"Chandrasekaran, D. et al. Generation of CAR-NK cells using a notch-mediated cell expansion and differentiation platform. Blood 136(Supplement 1), 42\u201342. https:\/\/doi.org\/10.1182\/BLOOD-2020-142813 (2020).","journal-title":"Blood"},{"issue":"19","key":"30732_CR21","doi-asserted-by":"publisher","first-page":"2452","DOI":"10.1182\/BLOODADVANCES.2018019059","volume":"2","author":"X Zhao","year":"2018","unstructured":"Zhao, X. et al. NK cell development in a human stem cell niche: KIR expression occurs independently of the presence of HLA class I ligands. Blood Adv. 2(19), 2452. https:\/\/doi.org\/10.1182\/BLOODADVANCES.2018019059 (2018).","journal-title":"Blood Adv."},{"issue":"4","key":"30732_CR22","doi-asserted-by":"publisher","first-page":"258","DOI":"10.1159\/000010327","volume":"50","author":"C Yap","year":"2000","unstructured":"Yap, C. et al. Variability in CD34+ cell counts in umbilical cord blood: Implications for cord blood transplants. Gynecol. Obstet. Invest. 50(4), 258\u2013259. https:\/\/doi.org\/10.1159\/000010327 (2000).","journal-title":"Gynecol. Obstet. Invest."},{"issue":"12","key":"30732_CR23","doi-asserted-by":"publisher","first-page":"2585","DOI":"10.1016\/J.STEMCR.2022.10.006","volume":"17","author":"M Jones","year":"2022","unstructured":"Jones, M., Cunningham, A., Frank, N. & Sethi, D. The monoculture of cord-blood-derived CD34+ cells by an automated, membrane-based dynamic perfusion system with a novel cytokine cocktail. Stem Cell Rep. 17(12), 2585. https:\/\/doi.org\/10.1016\/J.STEMCR.2022.10.006 (2022).","journal-title":"Stem Cell Rep."},{"key":"30732_CR24","doi-asserted-by":"crossref","unstructured":"Kogler, G. & Liedtke, S. Biology of umbilical cord blood cells. In Hematopoietic Stem Cell Transplantation and Cellular Therapies for Autoimmune Diseases. Published online January 1, 92\u2013101 (2021).","DOI":"10.1201\/9781315151366-11"},{"issue":"9","key":"30732_CR25","doi-asserted-by":"publisher","DOI":"10.1371\/JOURNAL.PONE.0012954","volume":"5","author":"M Ichii","year":"2010","unstructured":"Ichii, M. et al. The density of CD10 corresponds to commitment and progression in the human B lymphoid lineage. PLoS ONE 5(9), e12954. https:\/\/doi.org\/10.1371\/JOURNAL.PONE.0012954 (2010).","journal-title":"PLoS ONE"},{"issue":"13","key":"30732_CR26","doi-asserted-by":"publisher","first-page":"1059","DOI":"10.1182\/BLOOD.2019001289","volume":"134","author":"S O\u2019Byrne","year":"2019","unstructured":"O\u2019Byrne, S. et al. Discovery of a CD10-negative B-progenitor in human fetal life identifies unique ontogeny-related developmental programs. Blood 134(13), 1059\u20131071. https:\/\/doi.org\/10.1182\/BLOOD.2019001289 (2019).","journal-title":"Blood"},{"issue":"5822","key":"30732_CR27","doi-asserted-by":"publisher","first-page":"291","DOI":"10.1126\/SCIENCE.1139412","volume":"316","author":"AJ Bankovich","year":"2007","unstructured":"Bankovich, A. J. et al. Structural insight into pre-B cell receptor function. Science. (1979) 316(5822), 291\u2013294. https:\/\/doi.org\/10.1126\/SCIENCE.1139412 (2007).","journal-title":"Science. (1979)"},{"issue":"12","key":"30732_CR28","doi-asserted-by":"publisher","first-page":"2257","DOI":"10.1038\/SJ.LEU.2403202;KWRD","volume":"17","author":"JJM van Dongen","year":"2003","unstructured":"van Dongen, J. J. M. et al. Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: Report of the BIOMED-2 concerted action BMH4-CT98-3936. Leukemia 17(12), 2257\u20132317. https:\/\/doi.org\/10.1038\/SJ.LEU.2403202;KWRD (2003).","journal-title":"Leukemia"},{"key":"30732_CR29","doi-asserted-by":"publisher","DOI":"10.3389\/FIMMU.2021.610789\/XML\/NLM","volume":"12","author":"X Wang","year":"2021","unstructured":"Wang, X. & Zhao, X. Y. Transcription factors associated with IL-15 cytokine signaling during NK cell development. Front. Immunol. 12, 610789. https:\/\/doi.org\/10.3389\/FIMMU.2021.610789\/XML\/NLM (2021).","journal-title":"Front. Immunol."},{"issue":"1","key":"30732_CR30","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1084\/JEM.20082013","volume":"206","author":"ND Huntington","year":"2009","unstructured":"Huntington, N. D. et al. IL-15 trans-presentation promotes human NK cell development and differentiation in vivo. J. Exp. Med. 206(1), 25\u201334. https:\/\/doi.org\/10.1084\/JEM.20082013 (2009).","journal-title":"J. Exp. Med."},{"issue":"4","key":"30732_CR31","doi-asserted-by":"publisher","first-page":"310","DOI":"10.1038\/CMI.2012.17","volume":"9","author":"M Luevano","year":"2012","unstructured":"Luevano, M., Madrigal, A. & Saudemont, A. Generation of natural killer cells from hematopoietic stem cells in vitro for immunotherapy. Cell Mol. Immunol. 9(4), 310. https:\/\/doi.org\/10.1038\/CMI.2012.17 (2012).","journal-title":"Cell Mol. Immunol."},{"issue":"9","key":"30732_CR32","doi-asserted-by":"publisher","first-page":"703","DOI":"10.1038\/nri2154","volume":"7","author":"ND Huntington","year":"2007","unstructured":"Huntington, N. D., Vosshenrich, C. A. J. & Di Santo, J. P. Developmental pathways that generate natural-killer-cell diversity in mice and humans. Nat. Rev. Immunol. 7(9), 703\u2013714. https:\/\/doi.org\/10.1038\/nri2154 (2007).","journal-title":"Nat. Rev. Immunol."},{"issue":"126","key":"30732_CR33","doi-asserted-by":"publisher","first-page":"56191","DOI":"10.3791\/56191","volume":"2017","author":"F Kandarian","year":"2017","unstructured":"Kandarian, F., Sunga, G. M., Arango-Saenz, D. & Rossetti, M. A flow cytometry-based cytotoxicity assay for the assessment of human NK cell activity. J. Vis. Exp. 2017(126), 56191. https:\/\/doi.org\/10.3791\/56191 (2017).","journal-title":"J. Vis. Exp."},{"issue":"18","key":"30732_CR34","doi-asserted-by":"publisher","first-page":"5325","DOI":"10.1182\/BLOODADVANCES.2023009808","volume":"7","author":"MNE Sommarin","year":"2023","unstructured":"Sommarin, M. N. E. et al. Single-cell multiomics of human fetal hematopoiesis define a developmental-specific population and a fetal signature. Blood Adv. 7(18), 5325\u20135340. https:\/\/doi.org\/10.1182\/BLOODADVANCES.2023009808 (2023).","journal-title":"Blood Adv."},{"issue":"7778","key":"30732_CR35","doi-asserted-by":"publisher","first-page":"365","DOI":"10.1038\/S41586-019-1652-Y","volume":"574","author":"DM Popescu","year":"2019","unstructured":"Popescu, D. M. et al. Decoding human fetal liver haematopoiesis. Nature 574(7778), 365. https:\/\/doi.org\/10.1038\/S41586-019-1652-Y (2019).","journal-title":"Nature"},{"issue":"1","key":"30732_CR36","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/ncomms1475","volume":"2","author":"T Nojima","year":"2011","unstructured":"Nojima, T. et al. In-vitro derived germinal centre B cells differentially generate memory B or plasma cells in vivo. Nat. Commun. 2(1), 1\u201311. https:\/\/doi.org\/10.1038\/ncomms1475 (2011).","journal-title":"Nat. Commun."},{"issue":"12","key":"30732_CR37","doi-asserted-by":"publisher","DOI":"10.1371\/JOURNAL.PONE.0243769","volume":"15","author":"F Dubois","year":"2020","unstructured":"Dubois, F. et al. Toward a better definition of hematopoietic progenitors suitable for B cell differentiation. PLoS ONE 15(12), e0243769. https:\/\/doi.org\/10.1371\/JOURNAL.PONE.0243769 (2020).","journal-title":"PLoS ONE"},{"issue":"11","key":"30732_CR38","doi-asserted-by":"publisher","DOI":"10.1371\/JOURNAL.PONE.0050495","volume":"7","author":"J Calvo","year":"2012","unstructured":"Calvo, J., BenYoucef, A., Baijer, J., Rouyez, M. C. & Pflumio, F. Assessment of human multi-potent hematopoietic stem\/progenitor cell potential using a single in vitro screening system. PLoS ONE 7(11), e50495. https:\/\/doi.org\/10.1371\/JOURNAL.PONE.0050495 (2012).","journal-title":"PLoS ONE"},{"issue":"3","key":"30732_CR39","doi-asserted-by":"publisher","first-page":"362","DOI":"10.1016\/J.DEVCEL.2017.12.005","volume":"44","author":"C B\u00f6iers","year":"2018","unstructured":"B\u00f6iers, C. et al. A human IPS model implicates embryonic B-myeloid fate restriction as developmental susceptibility to B acute lymphoblastic leukemia-associated ETV6-RUNX1. Dev. Cell. 44(3), 362. https:\/\/doi.org\/10.1016\/J.DEVCEL.2017.12.005 (2018).","journal-title":"Dev. Cell."},{"issue":"1","key":"30732_CR40","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1084\/JEM.191.1.5","volume":"191","author":"R Carsetti","year":"2000","unstructured":"Carsetti, R. The development of B cells in the bone marrow is controlled by the balance between cell-autonomous mechanisms and signals from the microenvironment. J. Exp. Med. 191(1), 5. https:\/\/doi.org\/10.1084\/JEM.191.1.5 (2000).","journal-title":"J. Exp. Med."},{"issue":"1","key":"30732_CR41","doi-asserted-by":"publisher","first-page":"12","DOI":"10.1002\/STEM.5530100104","volume":"10","author":"K Dorshkind","year":"1992","unstructured":"Dorshkind, K. & Landreth, K. S. Regulation of B cell differentiation by bone marrow stromal cells. Stem Cells. 10(1), 12\u201317. https:\/\/doi.org\/10.1002\/STEM.5530100104 (1992).","journal-title":"Stem Cells."},{"issue":"8","key":"30732_CR42","doi-asserted-by":"publisher","first-page":"707","DOI":"10.1016\/J.EXPHEM.2014.06.008","volume":"42","author":"CE Schmitt","year":"2014","unstructured":"Schmitt, C. E., Lizama, C. O. & Zovein, A. C. From transplantation to transgenics: Mouse models of developmental hematopoiesis. Exp. Hematol. 42(8), 707\u2013716. https:\/\/doi.org\/10.1016\/J.EXPHEM.2014.06.008 (2014).","journal-title":"Exp. Hematol."},{"issue":"1","key":"30732_CR43","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/srep15680","volume":"5","author":"D Reichert","year":"2015","unstructured":"Reichert, D. et al. Phenotypic, morphological and adhesive differences of human hematopoietic progenitor cells cultured on murine versus human mesenchymal stromal cells. Sci. Rep. 5(1), 1\u201314 (2015).","journal-title":"Sci. Rep."},{"issue":"1","key":"30732_CR44","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41467-025-57096-y","volume":"16","author":"H Komic","year":"2025","unstructured":"Komic, H. et al. Continuous map of early hematopoietic stem cell differentiation across human lifetime. Nat. Commun. 16(1), 1\u201317. https:\/\/doi.org\/10.1038\/s41467-025-57096-y (2025).","journal-title":"Nat. Commun."},{"issue":"5","key":"30732_CR45","doi-asserted-by":"publisher","first-page":"587","DOI":"10.1016\/j.exphem.2007.12.013","volume":"36","author":"M Ichii","year":"2008","unstructured":"Ichii, M. et al. Regulation of human B lymphopoiesis by the transforming growth factor-\u03b2 superfamily in a newly established coculture system using human mesenchymal stem cells as a supportive microenvironment. Exp Hematol. 36(5), 587\u2013597. https:\/\/doi.org\/10.1016\/j.exphem.2007.12.013 (2008).","journal-title":"Exp Hematol."},{"issue":"1","key":"30732_CR46","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1016\/j.biologicals.2009.08.014","volume":"38","author":"JP Pilili","year":"2010","unstructured":"Pilili, J. P. et al. Testing genotoxicity and cytotoxicity strategies for the evaluation of commercial radiosterilized fetal calf sera. Biologicals 38(1), 135\u2013143. https:\/\/doi.org\/10.1016\/j.biologicals.2009.08.014 (2010).","journal-title":"Biologicals"},{"issue":"3","key":"30732_CR47","doi-asserted-by":"publisher","DOI":"10.1371\/JOURNAL.PONE.0212502","volume":"14","author":"B Mahadik","year":"2019","unstructured":"Mahadik, B., Hannon, B. & Harley, B. A. C. A computational model of feedback-mediated hematopoietic stem cell differentiation in vitro. PLoS ONE 14(3), e0212502. https:\/\/doi.org\/10.1371\/JOURNAL.PONE.0212502 (2019).","journal-title":"PLoS ONE"},{"issue":"12","key":"30732_CR48","doi-asserted-by":"publisher","DOI":"10.1371\/JOURNAL.PONE.0243769","volume":"15","author":"F Dubois","year":"2020","unstructured":"Dubois, F. et al. Toward a better definition of hematopoietic progenitors suitable for B cell differentiation. PLoS ONE 15(12), e0243769. https:\/\/doi.org\/10.1371\/JOURNAL.PONE.0243769 (2020).","journal-title":"PLoS ONE"},{"issue":"4","key":"30732_CR49","doi-asserted-by":"publisher","first-page":"410","DOI":"10.1046\/J.1365-2567.2001.01336.X","volume":"104","author":"Y Hirose","year":"2001","unstructured":"Hirose, Y. et al. B-cell precursors differentiated from cord blood CD34+ cells are more immature than those derived from granulocyte colony-stimulating factor-mobilized peripheral blood CD34+ cells. Immunology 104(4), 410. https:\/\/doi.org\/10.1046\/J.1365-2567.2001.01336.X (2001).","journal-title":"Immunology"},{"issue":"4","key":"30732_CR50","doi-asserted-by":"publisher","first-page":"421","DOI":"10.4252\/WJSC.V6.I4.421","volume":"6","author":"M Ichii","year":"2014","unstructured":"Ichii, M., Oritani, K. & Kanakura, Y. Early B lymphocyte development: Similarities and differences in human and mouse. World J. Stem Cells 6(4), 421\u2013431. https:\/\/doi.org\/10.4252\/WJSC.V6.I4.421 (2014).","journal-title":"World J. Stem Cells"},{"key":"30732_CR51","doi-asserted-by":"publisher","first-page":"7801781","DOI":"10.1155\/2016\/7801781","volume":"2016","author":"G Bonnaure","year":"2016","unstructured":"Bonnaure, G., Gervais-St-Amour, C. & N\u00e9ron, S. Bone marrow mesenchymal stem cells enhance the differentiation of human switched memory B lymphocytes into plasma cells in serum-free medium. J. Immunol. Res. 2016, 7801781. https:\/\/doi.org\/10.1155\/2016\/7801781 (2016).","journal-title":"J. Immunol. Res."},{"issue":"1","key":"30732_CR52","doi-asserted-by":"publisher","DOI":"10.1371\/JOURNAL.PONE.0087086","volume":"9","author":"M Luevano","year":"2014","unstructured":"Luevano, M. et al. Frozen cord blood hematopoietic stem cells differentiate into higher numbers of functional natural killer cells in vitro than mobilized hematopoietic stem cells or Freshly Isolated cord blood hematopoietic stem cells. PLoS ONE 9(1), e87086. https:\/\/doi.org\/10.1371\/JOURNAL.PONE.0087086 (2014).","journal-title":"PLoS ONE"},{"issue":"1","key":"30732_CR53","doi-asserted-by":"publisher","first-page":"295","DOI":"10.1186\/S13287-023-03461-X","volume":"14","author":"W Wen","year":"2023","unstructured":"Wen, W. et al. Enhancing cord blood stem cell-derived NK cell growth and differentiation through hyperosmosis. Stem Cell. Res. Ther. 14(1), 295. https:\/\/doi.org\/10.1186\/S13287-023-03461-X (2023).","journal-title":"Stem Cell. Res. Ther."},{"key":"30732_CR54","doi-asserted-by":"publisher","first-page":"1531736","DOI":"10.3389\/FIMMU.2025.1531736\/PDF","volume":"16","author":"M Martin Corredera","year":"2025","unstructured":"Martin Corredera, M. et al. Feeder-cell-free system for ex vivo production of natural killer cells from cord blood hematopoietic stem and progenitor cells. Front. Immunol. 16, 1531736. https:\/\/doi.org\/10.3389\/FIMMU.2025.1531736\/PDF (2025).","journal-title":"Front. Immunol."},{"issue":"1","key":"30732_CR55","doi-asserted-by":"publisher","first-page":"299","DOI":"10.4049\/JIMMUNOL.162.1.299","volume":"162","author":"KS Wang","year":"1999","unstructured":"Wang, K. S., Ritz, J. & Frank, D. A. IL-2 induces STAT4 activation in primary NK cells and NK cell lines, but not in T cells. J. Immunol. 162(1), 299\u2013304. https:\/\/doi.org\/10.4049\/JIMMUNOL.162.1.299 (1999).","journal-title":"J. Immunol."},{"key":"30732_CR56","doi-asserted-by":"publisher","first-page":"1531736","DOI":"10.3389\/FIMMU.2025.1531736\/XML\/NLM","volume":"16","author":"M Martin Corredera","year":"2025","unstructured":"Martin Corredera, M. et al. Feeder-cell-free system for ex vivo production of natural killer cells from cord blood hematopoietic stem and progenitor cells. Front. Immunol. 16, 1531736. https:\/\/doi.org\/10.3389\/FIMMU.2025.1531736\/XML\/NLM (2025).","journal-title":"Front. Immunol."},{"issue":"3","key":"30732_CR57","doi-asserted-by":"publisher","first-page":"1044","DOI":"10.4049\/JIMMUNOL.1301815","volume":"192","author":"H Kraus","year":"2014","unstructured":"Kraus, H. et al. A feeder-free differentiation system identifies autonomously proliferating B cell precursors in human bone marrow. J. Immunol. 192(3), 1044\u20131054. https:\/\/doi.org\/10.4049\/JIMMUNOL.1301815 (2014).","journal-title":"J. Immunol."},{"issue":"7","key":"30732_CR58","doi-asserted-by":"publisher","first-page":"1422","DOI":"10.1182\/BLOOD-2008-09-177139","volume":"113","author":"XM Luo","year":"2009","unstructured":"Luo, X. M. et al. Engineering human hematopoietic stem\/progenitor cells to produce a broadly neutralizing anti-HIV antibody after in vitro maturation to human B lymphocytes. Blood 113(7), 1422\u20131431. https:\/\/doi.org\/10.1182\/BLOOD-2008-09-177139 (2009).","journal-title":"Blood"},{"key":"30732_CR59","doi-asserted-by":"publisher","first-page":"1531736","DOI":"10.3389\/FIMMU.2025.1531736","volume":"16","author":"M Martin Corredera","year":"2025","unstructured":"Martin Corredera, M. et al. Feeder-cell-free system for ex vivo production of natural killer cells from cord blood hematopoietic stem and progenitor cells. Front. Immunol. 16, 1531736. https:\/\/doi.org\/10.3389\/FIMMU.2025.1531736 (2025).","journal-title":"Front. Immunol."},{"issue":"10","key":"30732_CR60","doi-asserted-by":"publisher","first-page":"939","DOI":"10.1016\/j.jcyt.2021.05.001","volume":"23","author":"KB Lupo","year":"2021","unstructured":"Lupo, K. B., Moon, J. I., Chambers, A. M. & Matosevic, S. Differentiation of natural killer cells from induced pluripotent stem cells under defined, serum- and feeder-free conditions. Cytotherapy. 23(10), 939\u2013952. https:\/\/doi.org\/10.1016\/j.jcyt.2021.05.001 (2021).","journal-title":"Cytotherapy."},{"issue":"4","key":"30732_CR61","doi-asserted-by":"publisher","first-page":"e70116","DOI":"10.1002\/HEM3.70116","volume":"9","author":"VH Jepsen","year":"2025","unstructured":"Jepsen, V. H. et al. H1\u20130 is a specific mediator of the repressive ETV6::RUNX1 transcriptional landscape in preleukemia and B cell acute lymphoblastic leukemia. Hemasphere. 9(4), e70116. https:\/\/doi.org\/10.1002\/HEM3.70116 (2025).","journal-title":"Hemasphere."},{"issue":"Supplement_1","key":"30732_CR62","doi-asserted-by":"publisher","first-page":"S26","DOI":"10.1093\/STCLTM\/SZAD047.025","volume":"12","author":"R Bhave","year":"2023","unstructured":"Bhave, R. et al. Abstract 24 development of an in vitro B-cell differentiation model for the characterization of the origin and causation of ETV6::RUNX1+ preleukemic cells. Stem Cells Transl. Med. 12(Supplement_1), S26\u2013S26. https:\/\/doi.org\/10.1093\/STCLTM\/SZAD047.025 (2023).","journal-title":"Stem Cells Transl. Med."},{"key":"30732_CR63","doi-asserted-by":"publisher","first-page":"S33","DOI":"10.1093\/stcltm\/szae062.030","volume":"13","author":"R Bhave","year":"2024","unstructured":"Bhave, R. et al. In-depth characterization of B-lineage populations arising during cord blood CD34+ HSPC differentiation for ETV6::RUNX1 progenitor identification. Stem Cells Transl. Med. 13, S33 (2024).","journal-title":"Stem Cells Transl. Med."},{"issue":"2","key":"30732_CR64","doi-asserted-by":"publisher","first-page":"158","DOI":"10.1093\/STMCLS\/SXAD085","volume":"42","author":"F Becker","year":"2024","unstructured":"Becker, F., Ouzin, M., Liedtke, S., Raba, K. & Kogler, G. DNA damage response after treatment of cycling and quiescent cord blood hematopoietic stem cells with distinct genotoxic noxae. Stem Cells. 42(2), 158\u2013171. https:\/\/doi.org\/10.1093\/STMCLS\/SXAD085 (2024).","journal-title":"Stem Cells."},{"issue":"3","key":"30732_CR65","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1089\/SCD.1.1996.5.213","volume":"5","author":"DR Sutherland","year":"1996","unstructured":"Sutherland, D. R., Anderson, L., Keeney, M., Nayar, R. & Chin-Yee, I. The ISHAGE guidelines for CD34+ cell determination by flow cytometry. J. Hematother. Stem Cell Res. 5(3), 213\u2013226. https:\/\/doi.org\/10.1089\/SCD.1.1996.5.213 (1996).","journal-title":"J. Hematother. Stem Cell Res."},{"issue":"24","key":"30732_CR66","doi-asserted-by":"publisher","first-page":"2134","DOI":"10.3390\/CELLS13242134\/S1","volume":"13","author":"M Ouzin","year":"2024","unstructured":"Ouzin, M., Wesselborg, S., Fritz, G. & Kogler, G. Evaluation of genotoxic effects of N-methyl-N-nitroso-urea and etoposide on the differentiation potential of MSCs from umbilical cord blood and bone marrow. Cells 13(24), 2134. https:\/\/doi.org\/10.3390\/CELLS13242134\/S1 (2024).","journal-title":"Cells"},{"issue":"1","key":"30732_CR67","doi-asserted-by":"publisher","first-page":"2395","DOI":"10.1038\/S41467-019-10291-0","volume":"10","author":"D Pellin","year":"2019","unstructured":"Pellin, D. et al. A comprehensive single cell transcriptional landscape of human hematopoietic progenitors. Nat. Commun. 10(1), 2395. https:\/\/doi.org\/10.1038\/S41467-019-10291-0 (2019).","journal-title":"Nat. Commun."},{"issue":"3","key":"30732_CR68","doi-asserted-by":"publisher","first-page":"1044","DOI":"10.4049\/JIMMUNOL.1301815","volume":"192","author":"H Kraus","year":"2014","unstructured":"Kraus, H. et al. A feeder-free differentiation system identifies autonomously proliferating B cell precursors in human bone marrow. J. Immunol. 192(3), 1044\u20131054. https:\/\/doi.org\/10.4049\/JIMMUNOL.1301815 (2014).","journal-title":"J. Immunol."},{"issue":"1\u20132","key":"30732_CR69","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1016\/J.JIM.2004.08.008","volume":"294","author":"G Alter","year":"2004","unstructured":"Alter, G., Malenfant, J. M. & Altfeld, M. CD107a as a functional marker for the identification of natural killer cell activity. J. Immunol. Methods. 294(1\u20132), 15\u201322. https:\/\/doi.org\/10.1016\/J.JIM.2004.08.008 (2004).","journal-title":"J. Immunol. Methods."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-30732-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-30732-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-30732-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,19]],"date-time":"2025-12-19T09:50:44Z","timestamp":1766137844000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-30732-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,18]]},"references-count":69,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["30732"],"URL":"https:\/\/doi.org\/10.1038\/s41598-025-30732-9","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,18]]},"assertion":[{"value":"18 July 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"26 November 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"18 December 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}},{"value":"CB units for research were obtained from donations collected by the Jos\u00e9 Carreras Cord Blood Bank D\u00fcsseldorf with the informed consent of the mothers and ethical approval (Ethic commission votes 2975, 5279, 2020-1144, 2020-9221). All CB units had a gestational age\u2009\u2265\u200936\u00a0weeks.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}}],"article-number":"44132"}}