{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T17:51:09Z","timestamp":1790704269256,"version":"4.1.0"},"reference-count":47,"publisher":"Oxford University Press (OUP)","issue":"5","license":[{"start":{"date-parts":[[2026,5,27]],"date-time":"2026-05-27T00:00:00Z","timestamp":1779840000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"Colossal Biosciences"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Primordial germ cells (PGCs) are critical tools for genome engineering and conservation in birds. Although culture systems for chicken PGCs have been well established for nearly two decades, efforts to propagate PGCs from other avian species have proved exceptionally challenging, limiting the broader application of artificial reproductive technologies in birds. Here we report the first successful derivation and long-term culture of PGCs from the rock dove, or common pigeon (Columba livia). Guided by transcriptomic profiling of PGCs, we developed a species-specific medium that supports PGC maintenance and expansion. We identify insulin signaling as a requirement for survival under the conditions tested, and demonstrate that inhibition of the retinoic acid receptor, in the presence of vitamin A, supports propagation. Supplementation with bone morphogenetic protein 4, leukemia inhibitory factor (LIF), glial cell line-derived neurotrophic factor, and pleiotrophin further enhances PGC proliferation. Cultured cells express canonical germline markers and migrate to the gonads following injection into both rock dove and chicken embryos, confirming functional migratory competency. These findings establish a platform for germline manipulation and biobanking in Columbidae, broadening the potential applicability of reproductive technologies to conservation efforts.<\/jats:p>","DOI":"10.1093\/jhered\/esag045","type":"journal-article","created":{"date-parts":[[2026,5,27]],"date-time":"2026-05-27T11:42:13Z","timestamp":1779882133000},"page":"1130-1143","source":"Crossref","is-referenced-by-count":3,"title":["Long-term rock dove (\n                    <i>Columba livia<\/i>\n                    ) primordial germ cell culture: A tool towards avian conservation"],"prefix":"10.1093","volume":"117","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-8031-3407","authenticated-orcid":false,"given":"Martin W","family":"Nicholson","sequence":"first","affiliation":[{"name":"Colossal Biosciences , Austin, TX ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucas","family":"Moreira","sequence":"additional","affiliation":[{"name":"Colossal Biosciences , Austin, TX ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emily A","family":"Boyle","sequence":"additional","affiliation":[{"name":"Colossal Biosciences , Austin, TX ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evan","family":"McCabe","sequence":"additional","affiliation":[{"name":"Colossal Biosciences , Austin, TX ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adrianna","family":"Soriano","sequence":"additional","affiliation":[{"name":"Colossal Biosciences , Austin, TX ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Malcolm","family":"McSwain","sequence":"additional","affiliation":[{"name":"Colossal Biosciences , Austin, TX ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William N","family":"Feist","sequence":"additional","affiliation":[{"name":"Colossal Biosciences , Austin, TX ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samantha K","family":"Nguyen","sequence":"additional","affiliation":[{"name":"Colossal Biosciences , Austin, TX ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaitlin","family":"Steiger","sequence":"additional","affiliation":[{"name":"Colossal Biosciences , Austin, TX ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolas","family":"Alexandre","sequence":"additional","affiliation":[{"name":"Colossal Biosciences , Austin, TX ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Savannah J","family":"Hoyt","sequence":"additional","affiliation":[{"name":"Institute for Systems Genomics, University of Connecticut , Storrs, CT ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8596-5335","authenticated-orcid":false,"given":"Nicole M","family":"Tillquist","sequence":"additional","affiliation":[{"name":"Institute for Systems Genomics, University of Connecticut , Storrs, CT ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1525-6821","authenticated-orcid":false,"given":"Rachel J","family":"O\u2019Neill","sequence":"additional","affiliation":[{"name":"Institute for Systems Genomics, University of Connecticut , Storrs, CT ,","place":["United States"]},{"name":"Department of Molecular and Cell Biology, University of Connecticut , Storrs, CT ,","place":["United States"]},{"name":"Department of Genetics and Genome Sciences, UConn Health , Farmington, CT ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kennosuke","family":"Ichikawa","sequence":"additional","affiliation":[{"name":"The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh , Midlothian ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mike J","family":"McGrew","sequence":"additional","affiliation":[{"name":"The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh , Midlothian ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olivier","family":"Fedrigo","sequence":"additional","affiliation":[{"name":"Colossal Biosciences , Austin, TX ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Keyte","sequence":"additional","affiliation":[{"name":"Colossal Biosciences , Austin, TX ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,5,27]]},"reference":[{"key":"2026090307275731600_ref1","doi-asserted-by":"publisher","first-page":"4809","DOI":"10.3390\/ijms26104809","article-title":"Significance of midkine signaling in women\u2019s cancers: novel biomarker and therapeutic target","volume":"26","author":"Aller","year":"2025","journal-title":"Int J Mol Sci"},{"key":"2026090307275731600_ref2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-021-91490-y","article-title":"Cultivation and characterization of primordial germ cells from blue layer hybrids (Araucana crossbreeds) and generation of germline chimeric chickens","volume":"11","author":"Altgilbers","year":"2021","journal-title":"Sci Rep"},{"key":"2026090307275731600_ref3","doi-asserted-by":"publisher","first-page":"14976","DOI":"10.1073\/pnas.0807297105","article-title":"Stra8 and its inducer, retinoic acid, regulate meiotic initiation in both spermatogenesis and oogenesis in mice","volume":"105","author":"Anderson","year":"2008","journal-title":"Proc Natl Acad Sci USA"},{"key":"2026090307275731600_ref4","doi-asserted-by":"publisher","first-page":"126","DOI":"10.1101\/gad.224503","article-title":"Multipotent cell lineages in early mouse development depend on SOX2 function","volume":"17","author":"Avilion","year":"2003","journal-title":"Genes Dev"},{"key":"2026090307275731600_ref5","doi-asserted-by":"publisher","first-page":"659","DOI":"10.1038\/s41467-020-20812-x","article-title":"Direct allele introgression into pure chicken breeds using sire dam surrogate (SDS) mating","volume":"12","author":"Ballantyne","year":"2021","journal-title":"Nat Commun"},{"key":"2026090307275731600_ref6","doi-asserted-by":"publisher","first-page":"38","DOI":"10.1038\/nbt.4314","article-title":"Dimensionality reduction for visualizing single-cell data using UMAP","volume":"37","author":"Becht","year":"2019","journal-title":"Nat Biotechnol"},{"key":"2026090307275731600_ref7","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1900\/RDS.2020.16.24","article-title":"Insulin-like growth factor and its therapeutic potential for diabetes complications\u2014mechanisms and metabolic links: a review","volume":"16","author":"Biadgo","year":"2020","journal-title":"Rev Diabet Stud"},{"key":"2026090307275731600_ref8","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1016\/j.ydbio.2024.08.006","article-title":"Pronounced early differentiation underlies zebra finch gonadal germ cell development","volume":"517","author":"Biegler","year":"2025","journal-title":"Dev Biol"},{"key":"2026090307275731600_ref9","doi-asserted-by":"publisher","first-page":"596","DOI":"10.1126\/science.1125691","article-title":"Retinoid signaling determines germ cell fate in mice","volume":"312","author":"Bowles","year":"2006","journal-title":"Science"},{"key":"2026090307275731600_ref10","doi-asserted-by":"publisher","first-page":"411","DOI":"10.1038\/nbt.4096","article-title":"Integrating single-cell transcriptomic data across different conditions, technologies, and species","volume":"36","author":"Butler","year":"2018","journal-title":"Nat Biotechnol"},{"key":"2026090307275731600_ref11","doi-asserted-by":"publisher","first-page":"e108693","DOI":"10.1371\/journal.pone.0108693","article-title":"Insulin resistance induced by hyperinsulinemia coincides with a persistent alteration at the insulin receptor tyrosine kinase domain","volume":"9","author":"Catalano","year":"2014","journal-title":"PLoS One"},{"key":"2026090307275731600_ref12","doi-asserted-by":"publisher","first-page":"e0200515","DOI":"10.1371\/journal.pone.0200515","article-title":"Three-dimensional culture of chicken primordial germ cells in defined media containing the functional polymer FP003","volume":"13","author":"Chen","year":"2018","journal-title":"PLoS One"},{"key":"2026090307275731600_ref13","doi-asserted-by":"publisher","first-page":"1557","DOI":"10.1038\/s41587-023-01685-z","article-title":"Microfluidics-free single-cell genomics with templated emulsification","volume":"41","author":"Clark","year":"2023","journal-title":"Nat Biotechnol"},{"key":"2026090307275731600_ref14","doi-asserted-by":"publisher","first-page":"87","DOI":"10.1002\/aja.1001250105","article-title":"Origin and early migration of primordial germ cells in the chick embryo","volume":"125","author":"Clawson","year":"1969","journal-title":"Am J Anat"},{"key":"2026090307275731600_ref15","doi-asserted-by":"publisher","first-page":"1146","DOI":"10.1242\/bio.20122592","article-title":"Chicken primordial germ cells use the anterior vitelline veins to enter the embryonic circulation","volume":"1","author":"De Melo Bernardo","year":"2012","journal-title":"Biol Open"},{"key":"2026090307275731600_ref16","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1038\/s42003-025-07715-7","article-title":"Propagation of goose primordial germ cells in vitro relies on FGF and BMP signalling pathways","volume":"8","author":"Doddamani","year":"2025","journal-title":"Commun Biol"},{"key":"2026090307275731600_ref17","doi-asserted-by":"publisher","first-page":"647","DOI":"10.1016\/S0092-8674(02)01135-2","article-title":"Guidance of primordial germ cell migration by the chemokine SDF-1","volume":"111","author":"Doitsidou","year":"2002","journal-title":"Cell"},{"key":"2026090307275731600_ref19","doi-asserted-by":"publisher","first-page":"784","DOI":"10.1016\/j.stemcr.2021.02.015","article-title":"Highly efficient genome modification of cultured primordial germ cells with lentiviral vectors to generate transgenic songbirds","volume":"16","author":"Gessara","year":"2021","journal-title":"Stem Cell Rep"},{"key":"2026090307275731600_ref20","doi-asserted-by":"publisher","first-page":"52","DOI":"10.1002\/9780470514637.ch5","article-title":"Primordial germ cell formation in birds","volume":"182","author":"Ginsburg","year":"1994","journal-title":"CIBA Found Symp"},{"key":"2026090307275731600_ref21","doi-asserted-by":"publisher","first-page":"296","DOI":"10.1186\/s13059-019-1874-1","article-title":"Normalization and variance stabilization of single-cell RNA-seq data","volume":"20","author":"Hafemeister","year":"2019","journal-title":"Genome Biol"},{"key":"2026090307275731600_ref22","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1002\/jmor.1050880104","article-title":"A series of normal stages in the development of the chick embryo","volume":"88","author":"Hamburger","year":"1951","journal-title":"J Morphol"},{"key":"2026090307275731600_ref23","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1186\/s40104-018-0234-4","article-title":"Primordial germ cell-mediated transgenesis and genome editing in birds","volume":"9","author":"Han","year":"2018","journal-title":"J Anim Sci Biotechnol"},{"key":"2026090307275731600_ref26","doi-asserted-by":"publisher","first-page":"e14591","DOI":"10.1111\/rda.14591","article-title":"Innovations in poultry reproduction using cryopreserved avian germ cells","volume":"59","author":"Ichikawa","year":"2024","journal-title":"Reprod Domest Anim"},{"key":"2026090307275731600_ref27","volume-title":"The IUCN Red List of Threatened Species (Version 2025-1)","author":"IUCN","year":"2025"},{"key":"2026090307275731600_ref28","doi-asserted-by":"publisher","first-page":"1127","DOI":"10.1038\/s42003-024-06775-5","article-title":"PGC-based cryobanking and regeneration through germline chimera mating","volume":"7","author":"Kinoshita","year":"2024","journal-title":"Commun Biol"},{"key":"2026090307275731600_ref29","doi-asserted-by":"publisher","first-page":"766","DOI":"10.1038\/nature04831","article-title":"Germline transmission of genetically modified PGCs","volume":"441","author":"Lavoir","year":"2006","journal-title":"Nature"},{"key":"2026090307275731600_ref30","doi-asserted-by":"publisher","first-page":"e35664","DOI":"10.1371\/journal.pone.0035664","article-title":"Interspecific germline transmission of cultured PGCs","volume":"7","author":"Lavoir","year":"2012","journal-title":"PLoS One"},{"key":"2026090307275731600_ref31","doi-asserted-by":"publisher","first-page":"101207","DOI":"10.1016\/j.psj.2021.101207","article-title":"Successful cryopreservation and regeneration of a Hungarian native chicken breed using primordial germ cells","volume":"100","author":"L\u00e1z\u00e1r","year":"2021","journal-title":"Poult Sci"},{"key":"2026090307275731600_ref32","doi-asserted-by":"publisher","first-page":"e76014","DOI":"10.7554\/eLife.76014","article-title":"Single-cell transcriptome reveals insights into zebrafish ovary development","volume":"11","author":"Liu","year":"2022","journal-title":"eLife"},{"key":"2026090307275731600_ref33","doi-asserted-by":"publisher","first-page":"258","DOI":"10.1016\/j.ydbio.2017.03.002","article-title":"BMP signaling in animal reproductive system development","volume":"427","author":"Lochab","year":"2017","journal-title":"Dev Biol"},{"key":"2026090307275731600_ref34","doi-asserted-by":"publisher","first-page":"673","DOI":"10.1093\/ps\/86.4.673","article-title":"Developmental changes of plasma insulin and IGFs in chick embryos","volume":"86","author":"Lu","year":"2007","journal-title":"Poult Sci"},{"key":"2026090307275731600_ref35","doi-asserted-by":"publisher","first-page":"269","DOI":"10.1038\/s41592-024-02552-8","article-title":"Orthology inference at scale with FastOMA","volume":"22","author":"Majidian","year":"2025","journal-title":"Nat Methods"},{"key":"2026090307275731600_ref37","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1093\/oxfordjournals.jbchem.a003231","article-title":"Midkine and pleiotrophin","volume":"132","author":"Muramatsu","year":"2002","journal-title":"J Biochem"},{"key":"2026090307275731600_ref39","doi-asserted-by":"publisher","first-page":"1597","DOI":"10.3390\/biology14111597","article-title":"In vitro culture of avian primordial germ cells","volume":"14","author":"Nayga","year":"2025","journal-title":"Biology"},{"key":"2026090307275731600_ref40","doi-asserted-by":"publisher","first-page":"543","DOI":"10.1038\/s44358-025-00065-6","article-title":"Genome engineering in biodiversity conservation","volume":"1","author":"Oosterhout","year":"2025","journal-title":"Nat Rev Biodivers"},{"key":"2026090307275731600_ref42","doi-asserted-by":"publisher","first-page":"274","DOI":"10.1002\/mrd.20821","article-title":"Production of quail (Coturnix japonica) germline chimeras derived from in vitro-cultured gonadal primordial germ cells","volume":"75","author":"","year":"2007","journal-title":"Mol Reprod Dev"},{"key":"2026090307275731600_ref44","doi-asserted-by":"publisher","first-page":"3482","DOI":"10.1210\/en.2011-1456","article-title":"Midkine Promotes Proliferation of Primordial Germ Cells by Inhibiting the Expression of the Deleted in Azoospermia-Like Gene","volume":"153","author":"","year":"2012","journal-title":"Endocrinology"},{"key":"2026090307275731600_ref46","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1002\/aja.1000150404","article-title":"Origin and early history of the primordial germ-cells in the chick","volume":"15","author":"Swift","year":"1914","journal-title":"Am J Anat"},{"key":"2026090307275731600_ref47","doi-asserted-by":"publisher","first-page":"2206","DOI":"10.1071\/RD16289","article-title":"Successful chimera production in Hungarian goose","volume":"29","author":"","year":"2017","journal-title":"Reprod Fertil Dev"},{"key":"2026090307275731600_ref48","doi-asserted-by":"publisher","first-page":"509","DOI":"10.1016\/0093-691X(93)90404-S","article-title":"Production of germ line chimera by transfer of primordial germ cells in the domestic chicken (Gallusdomesticus)","volume":"40","author":"","year":"1993","journal-title":"Theriogenology"},{"key":"2026090307275731600_ref49","doi-asserted-by":"publisher","first-page":"928","DOI":"10.1242\/dev.145367","article-title":"Efficient TALEN-mediated gene targeting of chicken PGCs","volume":"144","author":"Taylor","year":"2017","journal-title":"Development"},{"key":"2026090307275731600_ref50","doi-asserted-by":"publisher","first-page":"145","DOI":"10.1095\/biolreprod.112.106526","article-title":"Minimal Concentrations of Retinoic Acid Induce Stimulation by Retinoic Acid 8 and Promote Entry into Meiosis in Isolated Pregonadal and Gonadal Mouse Primordial Germ Cells","volume":"88","author":"","year":"2013","journal-title":"Biol Reprod"},{"key":"2026090307275731600_ref51","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1017\/S0030605307001016","article-title":"Geographical patterns of threat among pigeons and doves (Columbidae)","volume":"41","author":"Walker","year":"2007","journal-title":"Oryx"},{"key":"2026090307275731600_ref52","doi-asserted-by":"publisher","first-page":"80","DOI":"10.1186\/1465-9921-11-80","volume":"11","author":"Weng","year":"2010","journal-title":"Respir Res"},{"key":"2026090307275731600_ref53","doi-asserted-by":"publisher","first-page":"1171","DOI":"10.1016\/j.stemcr.2015.10.008","article-title":"FGF, Insulin, and SMAD Signaling Cooperate for Avian Primordial Germ Cell Self-Renewal","volume":"5","author":"Whyte","year":"2015","journal-title":"Stem Cell Rep"},{"key":"2026090307275731600_ref55","doi-asserted-by":"publisher","DOI":"10.3389\/fphys.2021.562817","article-title":"Single-cell RNA sequencing analysis of chicken anterior pituitary: a bird\u2019s-eye view on vertebrate pituitary","volume":"12","author":"Zhang","year":"2021","journal-title":"Front Physiol"},{"key":"2026090307275731600_ref56","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1016\/j.jare.2024.01.012","article-title":"Single-cell RNA sequencing reveals oocyte\u2013granulosa crosstalk and regulatory networks driving chicken primordial follicle assembly","volume":"70","author":"Zhang","year":"2025","journal-title":"J Adv Res"}],"container-title":["Journal of Heredity"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/jhered\/advance-article-pdf\/doi\/10.1093\/jhered\/esag045\/68410712\/esag045.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jhered\/article-pdf\/117\/5\/1130\/68410712\/esag045.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jhered\/article-pdf\/117\/5\/1130\/68410712\/esag045.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T11:28:03Z","timestamp":1788434883000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jhered\/article\/117\/5\/1130\/8696112"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,27]]},"references-count":47,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,5,27]]},"published-print":{"date-parts":[[2026,9,3]]}},"URL":"https:\/\/doi.org\/10.1093\/jhered\/esag045","relation":{},"ISSN":["0022-1503","1465-7333"],"issn-type":[{"value":"0022-1503","type":"print"},{"value":"1465-7333","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2026,9]]},"published":{"date-parts":[[2026,5,27]]}}}