{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,29]],"date-time":"2026-10-29T12:16:42Z","timestamp":1793276202413,"version":"4.5.0"},"reference-count":54,"publisher":"SAGE Publications","issue":"8","license":[{"start":{"date-parts":[[2023,8,1]],"date-time":"2023-08-01T00:00:00Z","timestamp":1690848000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2023,8,1]],"date-time":"2023-08-01T00:00:00Z","timestamp":1690848000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Tissue Engineering Part C: Methods"],"published-print":{"date-parts":[[2023,8,1]]},"abstract":"<jats:p>\n                    Corneal nerves originate from the ophthalmic branch of the trigeminal nerve, which enters the cornea at the limbus radially from all directions toward the central cornea. The cell bodies of the sensory neurons of trigeminal nerve are located in the trigeminal ganglion (TG), while the axons are extended into the three divisions, including ophthalmic branch that supplies corneal nerves. Study of primary neuronal cultures established from the TG fibers can therefore provide a knowledge basis for corneal nerve biology and potentially be developed as an\n                    <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                    platform for drug testing. However, setting up primary neuron cultures from animal TG has been dubious with inconsistency among laboratories due to a lack of efficient isolation protocol, resulting in low yield and heterogenous cultures. In this study, we used a combined enzymatic digestion with collagenase and TrypLE to dissociate mouse TG while preserving nerve cell viability. A subsequent discontinuous Percoll density gradient followed by mitotic inhibitor treatment effectively diminished the contamination of non-neuronal cells. Using this method, we reproducibly generated high yield and homogenous primary TG neuron cultures. Similar efficiency of nerve cell isolation and culture was further obtained for TG tissue cryopreserved for short (1 week) and long duration (3 months), compared to freshly isolated tissues. In conclusion, this optimized protocol shows a promising potential to standardize TG nerve culture and generate a high-quality corneal nerve model for drug testing and neurotoxicity studies.\n                  <\/jats:p>","DOI":"10.1089\/ten.tec.2023.0054","type":"journal-article","created":{"date-parts":[[2023,5,22]],"date-time":"2023-05-22T06:03:36Z","timestamp":1684735416000},"page":"381-393","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":15,"title":["Culture of Primary Neurons from Dissociated and Cryopreserved Mouse Trigeminal Ganglion"],"prefix":"10.1177","volume":"29","author":[{"given":"Molly Tzu-Yu","family":"Lin","sequence":"first","affiliation":[{"name":"Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore, Singapore."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabelle Xin Yu","family":"Lee","sequence":"additional","affiliation":[{"name":"Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore, Singapore."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei-Li","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan."},{"name":"Advanced Ocular Surface and Corneal Nerve Research Center, National Taiwan University, Taipei, Taiwan."},{"name":"Department of Ophthalmology, College of Medicine, National Taiwan University, Taipei, Taiwan."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mei-Yun","family":"Chen","sequence":"additional","affiliation":[{"name":"Advanced Ocular Surface and Corneal Nerve Research Center, National Taiwan University, Taipei, Taiwan."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jodhbir S.","family":"Mehta","sequence":"additional","affiliation":[{"name":"Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore, Singapore."},{"name":"Corneal and External Eye Disease Department, Singapore National Eye Centre, Singapore, Singapore."},{"name":"Ophthalmology and Visual Sciences Academic Clinical Program, Duke-NUS Medical School, Singapore, Singapore."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gary H. 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