{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T02:07:23Z","timestamp":1782871643445,"version":"3.54.5"},"reference-count":46,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2025,10,23]],"date-time":"2025-10-23T00:00:00Z","timestamp":1761177600000},"content-version":"vor","delay-in-days":22,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2025,10,1]],"date-time":"2025-10-01T00:00:00Z","timestamp":1759276800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2025,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Prostate diseases are increasing globally, necessitating an appropriate model system for investigation. Ex vivo organ models help bridge the gap between organoids, assembloids, and in vivo models. This article represents a robust and reproducible approach for maintaining a functional ex vivo whole\u2010organ prostate gland from the earthworm\n                    <jats:italic>Eudrilus eugeniae<\/jats:italic>\n                    , providing a unique platform for investigating organ\u2010level regeneration, drug screening, and organoid\u2010based research. Sexually mature\n                    <jats:italic>E. eugeniae<\/jats:italic>\n                    worms, which possess a pair of euprostate glands, are pre\u2010processed, meticulously dissected in a sterile environment, and surface sterilized. The glands are treated sequentially in a series of buffers and antibiotic solutions and, without exposure to osmotic stress, are transferred to Leibovitz's L\u201015 medium containing 10% FBS. The ex vivo prostate glands are incubated at 23\u00b0C, simulating the organism's normal temperature. The viability and stress response of the gland are assessed through the rhythmic activity and frequency of prostatic fluids and cells discharged from the anterior duct. Microbial contamination is reduced through early detection and antibiotic treatment. Amputation of the posterior lobe facilitates regeneration via wound closure, contraction, and tissue remodeling, which are enhanced by the use of valproic acid. Released prostatic fluid facilitates noninvasive molecular diagnosis and monitoring of therapeutic response. The cell aggregates released into the medium from the posterior lobe form spheroids, which subsequently grow into organoids and mini\u2010organ structures through self\u2010assembly and cellular bridging. Overall, the developed ex vivo prostate model meets 3R principles for minimizing animal experimentation. \u00a9 2025 Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Pre\u2010processing, dissection, and processing of prostate gland for ex vivo maintenance\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol<\/jats:bold>\n                    : Assessing ex vivo viability of prostate gland and handling contamination\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : Functional ex vivo prostate gland regeneration, drug screening, and spheroid, organoid, and mini\u2010organ formation\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70234","type":"journal-article","created":{"date-parts":[[2025,10,23]],"date-time":"2025-10-23T11:18:12Z","timestamp":1761218292000},"source":"Crossref","is-referenced-by-count":1,"title":["Long\u2010Term Ex Vivo Maintenance of Functional Whole Prostate Glands from the Earthworm\n                    <i>Eudrilus eugeniae<\/i>\n                    and Its Applications in Regeneration, Drug Screening, and 3D Models"],"prefix":"10.1002","volume":"5","author":[{"given":"Karthikeyan","family":"Subbiahanadar Chelladurai","sequence":"first","affiliation":[{"name":"Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre Sathyabama Institute of Science and Technology (Deemed to be University)  Chennai Tamil Nadu India"},{"name":"School of Health Sciences Purdue University  West Lafayette Indiana"},{"name":"These authors contributed equally to the work"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nandha Kumar","family":"Suresh","sequence":"additional","affiliation":[{"name":"Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre Sathyabama Institute of Science and Technology (Deemed to be University)  Chennai Tamil Nadu India"},{"name":"These authors contributed equally to the work"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0363-0529","authenticated-orcid":false,"given":"Jackson Durairaj","family":"Selvan Christyraj","sequence":"additional","affiliation":[{"name":"Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre Sathyabama Institute of Science and Technology (Deemed to be University)  Chennai Tamil Nadu India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kamarajan","family":"Rajagopalan","sequence":"additional","affiliation":[{"name":"Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre Sathyabama Institute of Science and Technology (Deemed to be University)  Chennai Tamil Nadu India"},{"name":"Present address: Vascular Biology Center, Augusta University  Augusta Georgia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johnson Retnaraj Samuel","family":"Selvan Christyraj","sequence":"additional","affiliation":[{"name":"Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre Sathyabama Institute of Science and Technology (Deemed to be University)  Chennai Tamil Nadu India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Puja","family":"Das","sequence":"additional","affiliation":[{"name":"Molecular 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