{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,20]],"date-time":"2025-09-20T18:44:06Z","timestamp":1758393846457,"version":"3.40.5"},"reference-count":24,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2021,3,3]],"date-time":"2021-03-03T00:00:00Z","timestamp":1614729600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["KFO296"],"award-info":[{"award-number":["KFO296"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2021,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>This panel was designed for the identification and detailed characterization of the different developmental steps of human thymocytes. We optimized the panel for fresh tissue in order to provide an unbiased analysis of T cell development. Accurate selection of antibodies and precise gating allow us to phenotype 14 major stages of human thymocyte development and illustrate the trajectories of T cell development from early thymic progenitors (ETP) to mature T cells that are ready to populate the periphery. The panel identifies ETPs, T\u2010lineage\u2010committed cells (TC), CD34\u2010positive immature single\u2010positive CD4 cells (ISP4 CD34+), CD34\u2010negative immature single\u2010positive CD4 cells (ISP4 CD34\u2010), CD45\u2010low early double\u2010positive cells (EDP CD45low), CD45\u2010high early double\u2010positive cells (EDP CD45high), late double\u2010positive cells (LDP), single\u2010positive CD4 cells (SP4), single\u2010positive CD8 cells (SP8), ready\u2010to\u2010egress single\u2010positive CD4 cells (rSP4), ready\u2010to\u2010egress single\u2010positive CD8 cells (rSP8), T \u03b3\u03b4 cells (T\u03b3\u03b4), T regulatory cells (Treg), and ready\u2010to\u2010egress T regulatory cells (rTreg). To highlight important checkpoints during T cell development, we added antibodies relevant for specific developmental steps to the panel. These include CD1a to define TCs, CD28 as a marker for \u00df\u2010selection and CD69 in combination with CD45RA to determine the maturation stage of thymocytes shortly before they become ready to egress the thymus and colonize the periphery. Moreover, Annexin V, as a marker for apoptosis, provides valuable extra information concerning the apoptotic death of thymocytes. Currently, we use this panel to identify aberrations in T cell development in health and disease.<\/jats:p>","DOI":"10.1002\/cyto.a.24326","type":"journal-article","created":{"date-parts":[[2021,3,3]],"date-time":"2021-03-03T09:19:36Z","timestamp":1614763176000},"page":"875-879","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["OMIP 073: Analysis of human thymocyte development with\u00a0a\u00a014\u2010color flow cytometry panel"],"prefix":"10.1002","volume":"99","author":[{"given":"Sarah\u2010Jolan","family":"Bremer","sequence":"first","affiliation":[{"name":"Department of Immunology University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura","family":"Glau","sequence":"additional","affiliation":[{"name":"Department of Immunology University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christina","family":"Gehbauer","sequence":"additional","affiliation":[{"name":"Department of Immunology University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annika","family":"Boxnick","sequence":"additional","affiliation":[{"name":"Department of Immunology University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel","family":"Biermann","sequence":"additional","affiliation":[{"name":"Surgery for Congenital Heart Disease, University Heart &amp; Vascular Center Hamburg University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J\u00f6rg Siegmar","family":"Sachweh","sequence":"additional","affiliation":[{"name":"Surgery for Congenital Heart Disease, University Heart &amp; Vascular Center Hamburg University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eva","family":"Tolosa","sequence":"additional","affiliation":[{"name":"Department of Immunology University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5147-2281","authenticated-orcid":false,"given":"Anna","family":"Gieras","sequence":"additional","affiliation":[{"name":"Department of Immunology University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2021,3,3]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nri913"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00281-008-0135-2"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/nri1257"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/nri798"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2020.03.019"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aay3224"},{"issue":"2434","key":"e_1_2_7_8_1","first-page":"1","article-title":"CD maps\u2013dynamic profiling of CD1\u2013CD100 surface expression on human leukocyte and lymphocyte subsets","volume":"10","author":"Kalina T","year":"2019","journal-title":"Front Immunol"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cels.2019.09.008"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2020.10.024"},{"key":"e_1_2_7_11_1","first-page":"953","article-title":"Quantitation of apoptosis and necrosis by annexin v binding, propidium iodide uptake and flow cytometry","volume":"11","author":"Crowley LC","year":"2016","journal-title":"Cold Spring Harb Protoc"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-030-12040-5_4"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(02)00474-0"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2006.01.006"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-065X.1998.tb01231.x"},{"key":"e_1_2_7_16_1","first-page":"221","volume-title":"T\u2010cell development. 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