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The development of T cells is dependent on small numbers of uncommitted precursors that continually seed the thymus from the bone marrow. As they progress along the developmental pathway, there is a massive expansion in cell number to generate the necessary diversity in T\u2010cell receptor chain usage. It is recognized that there are two proliferative bursts that occur early in T\u2010cell development, one prior to \u03b2\u2010selection and one after, and these are responsible for the expansion. While the proliferation following \u03b2\u2010selection is well\u2010characterized, the earlier proliferative burst has yet to be precisely defined. In this study, we employ single\u2010cell RNA sequencing coupled to trajectory inference methods to pinpoint when in T\u2010cell development thymocytes are induced into cell cycle. We show that the first proliferative burst is initiated in the double\u2010negative (DN) 2a stage before T lineage commitment occurs, with cell cycling downregulated by the DN3a stage. A second burst is then initiated at the DN3b stage, immediately after \u03b2\u2010selection. We subsequently employ fluorescence\u2010activated cell sorting\u2013based analysis for DNA content to confirm these two proliferative bursts.<\/jats:p>","DOI":"10.1111\/imcb.12670","type":"journal-article","created":{"date-parts":[[2023,7,19]],"date-time":"2023-07-19T10:21:23Z","timestamp":1689762083000},"page":"766-774","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Mapping the two distinct proliferative bursts early in T\u2010cell development"],"prefix":"10.1111","volume":"101","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4923-082X","authenticated-orcid":false,"given":"Seungyoul","family":"Oh","sequence":"first","affiliation":[{"name":"St Vincent's Institute of Medical Research  Fitzroy VIC Australia"},{"name":"Department of Medicine (St Vincent's) University of Melbourne  Fitzroy VIC Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dhruti","family":"Parikh","sequence":"additional","affiliation":[{"name":"St Vincent's Institute of Medical Research  Fitzroy VIC Australia"},{"name":"Department of Medicine (St Vincent's) University of Melbourne  Fitzroy VIC Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiyao","family":"Xiao","sequence":"additional","affiliation":[{"name":"St Vincent's Institute of Medical Research  Fitzroy VIC Australia"},{"name":"Faculty of Science University of Melbourne  Parkville VIC Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Liu","sequence":"additional","affiliation":[{"name":"St Vincent's Institute of Medical Research  Fitzroy VIC Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karen","family":"Gu","sequence":"additional","affiliation":[{"name":"St Vincent's Institute of Medical Research  Fitzroy VIC Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3701-7397","authenticated-orcid":false,"given":"Mark MW","family":"Chong","sequence":"additional","affiliation":[{"name":"St Vincent's Institute of Medical Research  Fitzroy VIC Australia"},{"name":"Department of Medicine (St Vincent's) University of Melbourne  Fitzroy VIC Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,7,19]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.150.10.4244"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-020-00426-6"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1106652"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2006.05.010"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1000679"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/ni927"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/ni.1887"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2007.05.009"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(00)80329-5"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2011-12-395319"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2012.08.020"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1101\/gad.10.8.948"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2016-05-716480"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cels.2019.03.003"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4402"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-0969-x"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12864-018-4772-0"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2004.05.004"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.185.9.1549"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.716661"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1158\/2159-8290.CD-19-0471"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2006-02-005900"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1093\/intimm\/dxq020"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1535-6108(03)00301-5"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-019-0469-z"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.05.031"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-019-1874-1"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2021.04.048"},{"key":"e_1_2_8_30_1","unstructured":"TenenbaumD MaintainerB.KEGGREST: client\u2010side REST access to the Kyoto encyclopedia of genes and genomes (KEGG). 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