{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T12:14:54Z","timestamp":1753877694491,"version":"3.41.2"},"reference-count":19,"publisher":"Oxford University Press (OUP)","issue":"1","license":[{"start":{"date-parts":[[2022,11,23]],"date-time":"2022-11-23T00:00:00Z","timestamp":1669161600000},"content-version":"vor","delay-in-days":326,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Borne"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,6,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>During pregnancy, interactions between uterine immune cells and cells of the surrounding reproductive tissues are thought to be vital for regulating labour. The mechanism that specifically initiates spontaneous labour has not been determined, but distinct changes in uterine immune cell populations and their activation status have been observed during labour at term gestation. To understand the regulation of human labour by the immune system, the ability to isolate both immune cells and non-immune cells from the uterus is required. Here, we describe protocols developed in our laboratory to isolate single cells from uterine tissues, which preserve both immune and non-immune cell populations for further analysis. We provide detailed methods for isolating immune and non-immune cells from human myometrium, chorion, amnion and decidua, together with representative flow cytometry analysis of isolated cell populations present. The protocols can be completed in tandem and take approximately 4\u20135 h, resulting in single-cell suspensions that contain viable leucocytes, and non-immune cells in sufficient numbers for single-cell analysis approaches such as flow cytometry and single cell RNA sequencing (scRNAseq).<\/jats:p>","DOI":"10.1093\/oxfimm\/iqac010","type":"journal-article","created":{"date-parts":[[2022,11,23]],"date-time":"2022-11-23T10:10:49Z","timestamp":1669198249000},"source":"Crossref","is-referenced-by-count":2,"title":["Isolation of single cells from human uterus in the third trimester of pregnancy: myometrium, decidua, amnion and chorion"],"prefix":"10.1093","volume":"3","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7162-0774","authenticated-orcid":false,"given":"Alexander T H","family":"Cocker","sequence":"first","affiliation":[{"name":"Department of Metabolism, Digestion and Reproduction, Imperial College London , London, UK"},{"name":"Department of Structural Biology, Stanford University , Stanford, CA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emily M","family":"Whettlock","sequence":"additional","affiliation":[{"name":"Department of Metabolism, Digestion and Reproduction, Imperial College London , London, UK"},{"name":"Department of Infectious Disease, Imperial College London , London, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brendan","family":"Browne","sequence":"additional","affiliation":[{"name":"Department of Metabolism, Digestion and Reproduction, Imperial College London , London, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pei F","family":"Lai","sequence":"additional","affiliation":[{"name":"Department of Metabolism, Digestion and Reproduction, Imperial College London , London, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jonathan K H","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Metabolism, Digestion and Reproduction, Imperial College London , London, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sivatharjini P","family":"Sivarajasingam","sequence":"additional","affiliation":[{"name":"Department of Metabolism, Digestion and Reproduction, Imperial College London , London, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nesrina","family":"Imami","sequence":"additional","affiliation":[{"name":"Department of Infectious Disease, Imperial College London , London, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark R","family":"Johnson","sequence":"additional","affiliation":[{"name":"Department of Metabolism, Digestion and Reproduction, Imperial College London , London, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5654-5083","authenticated-orcid":false,"given":"Victoria","family":"Male","sequence":"additional","affiliation":[{"name":"Department of Metabolism, Digestion and Reproduction, Imperial College London , London, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2022,11,23]]},"reference":[{"key":"2023011714432298400_iqac010-B1","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1097\/00003081-198809000-00005","article-title":"Biomolecular processes in the initiation of parturition: Decidual activation","volume":"31","author":"Casey Linette","year":"1988","journal-title":"Clin Obstet Gynecol"},{"key":"2023011714432298400_iqac010-B2","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1095\/biolreprod.111.095505","article-title":"Macrophages infiltrate the human and rat decidua during term and preterm labor: Evidence that decidual inflammation precedes labor","volume":"86","author":"Hamilton","year":"2012","journal-title":"Biol Reprod"},{"key":"2023011714432298400_iqac010-B3","doi-asserted-by":"crossref","first-page":"957","DOI":"10.1084\/jem.20041214","article-title":"Combinations of maternal KIR and fetal HLA-C genes influence the risk of preeclampsia and reproductive success","volume":"200","author":"Hiby","year":"2004","journal-title":"J Exp Med"},{"key":"2023011714432298400_iqac010-B4","doi-asserted-by":"crossref","first-page":"4102","DOI":"10.1172\/JCI43998","article-title":"Maternal activating KIRs protect against human reproductive failure mediated by fetal HLA-C2","volume":"120","author":"Hiby","year":"2010","journal-title":"J Clin Invest"},{"key":"2023011714432298400_iqac010-B5","doi-asserted-by":"crossref","first-page":"4292","DOI":"10.4049\/jimmunol.1601279","article-title":"Activating KIR2DS4 is expressed by uterine NK cells and contributes to successful pregnancy","volume":"197","author":"Kennedy","year":"2016","journal-title":"J Immunol"},{"key":"2023011714432298400_iqac010-B6","doi-asserted-by":"crossref","first-page":"4264","DOI":"10.1172\/JCI68991","article-title":"Maternal uterine NK cell-activating receptor KIR2DS1 enhances placentation","volume":"123","author":"Xiong","year":"2013","journal-title":"J Clin Invest"},{"issue":"Unit 7","key":"2023011714432298400_iqac010-B7","first-page":"41","article-title":"Isolation of cells from the feto-maternal interface","volume":"Chapter 7","author":"Male","year":"2012","journal-title":"Curr Protoc Immunol"},{"key":"2023011714432298400_iqac010-B8","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1038\/s41467-019-14123-z","article-title":"Distinctive phenotypes and functions of innate lymphoid cells in human decidua during early pregnancy","volume":"11","author":"Huhn","year":"2020","journal-title":"Nat Commun"},{"key":"2023011714432298400_iqac010-B9","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1038\/s41586-018-0698-6","article-title":"Single-cell reconstruction of the early maternal-fetal interface in humans","volume":"563","author":"Vento-Tormo","year":"2018","journal-title":"Nature"},{"key":"2023011714432298400_iqac010-B10","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1111\/aji.12185","article-title":"Quantification of the predominant immune cell populations in decidua throughout human pregnancy","volume":"71","author":"Bartmann","year":"2014","journal-title":"Am J Reprod Immunol"},{"key":"2023011714432298400_iqac010-B11","doi-asserted-by":"crossref","first-page":"880438","DOI":"10.3389\/fimmu.2022.880438","article-title":"Dynamic changes in uterine NK cell subset frequency and function over the menstrual cycle and pregnancy","volume":"13","author":"Whettlock","year":"2022","journal-title":"Front Immunol"},{"key":"2023011714432298400_iqac010-B12","doi-asserted-by":"crossref","first-page":"1031","DOI":"10.1093\/biolre\/ioz144","article-title":"Distinct preterm labor phenotypes have unique inflammatory signatures and contraction associated protein profiles\u2020","volume":"101","author":"Singh","year":"2019","journal-title":"Biol Reprod"},{"key":"2023011714432298400_iqac010-B13","doi-asserted-by":"crossref","first-page":"3128010","DOI":"10.1155\/2019\/3128010","article-title":"Exhausted and senescent T cells at the maternal-fetal interface in preterm and term labor","volume":"2019","author":"Slutsky","year":"2019","journal-title":"J Immunol Res"},{"key":"2023011714432298400_iqac010-B14","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1095\/biolreprod66.2.445","article-title":"Immunolocalization of proinflammatory cytokines in myometrium, cervix, and fetal membranes during human parturition at term","volume":"66","author":"Young","year":"2002","journal-title":"Biol Reprod"},{"key":"2023011714432298400_iqac010-B15","doi-asserted-by":"crossref","DOI":"10.7554\/eLife.52004","article-title":"Single cell transcriptional signatures of the human placenta in term and preterm parturition","volume":"8","author":"Pique-Regi","year":"2019","journal-title":"eLife"},{"key":"2023011714432298400_iqac010-B16","article-title":"Uterine overdistention induces preterm labor mediated by inflammation: observations in pregnant women and nonhuman primates","volume":"213","author":"Waldorf","year":"2015","journal-title":"Am J Obstet Gynecol"},{"key":"2023011714432298400_iqac010-B17","doi-asserted-by":"crossref","first-page":"185","DOI":"10.3389\/fgene.2019.00185","article-title":"Myometrial transcriptional signatures of human parturition","volume":"10","author":"Stanfield","year":"2019","journal-title":"Front Genet"},{"key":"2023011714432298400_iqac010-B18","doi-asserted-by":"crossref","DOI":"10.1111\/aji.12776","article-title":"Gestational tissue inflammatory biomarkers at term labor: A systematic review of literature","volume":"79","author":"Hadley","year":"2018","journal-title":"Am J Reprod 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