{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,14]],"date-time":"2025-05-14T00:35:40Z","timestamp":1747182940203,"version":"3.40.5"},"reference-count":37,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2022,7,11]],"date-time":"2022-07-11T00:00:00Z","timestamp":1657497600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100008530","name":"European Regional Development Fund","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100008530","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Anat Histol Embryol"],"published-print":{"date-parts":[[2022,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Cartilage differentiates in rat limb buds cultivated in a chemically defined protein\u2010free medium in the same manner as in the richer serum\u2010supplemented medium. We aimed to investigate the remaining differentiation potential of pre\u2010cultivated limb buds by subsequent transplantation in vivo. Rat front (FLBs) and hind\u2010limb buds (HLBs) were isolated from Fischer rat dams at the 14th gestation day (GD 14) and cultivated at the air\u2010liquid interface in Eagle's Minimum Essential Medium (MEM) alone; with 5\u00a0\u03bcM of 5\u2010azacytidine (5azaC) or with rat serum (1:1). Overall growth was measured seven times during the culture by an ocular micrometre. After 14\u2009days, explants were transplanted under the kidney capsule of adult males. Growth of limb buds was significantly lower in all limb buds cultivated in MEM than in those cultivated with serum. In MEM with 5azaC, growth of LBs was significantly lower only on day 3 of culture. Afterwards, it was higher throughout the culture period, although a statistically significant difference was assessed only for HLBs. In transplants, mixed structures developed with the differentiated transmembranous bone, cartilage with enchondral ossification, bone\u2010marrow, sebaceous gland, and hair that have never been found in vitro. Nerves differentiated only in transplants precultivated in the serum\u2010supplemented medium. We conclude that pre\u2010cultivation of LBs in a chemically defined protein\u2010free medium does not restrict osteogenesis and formation of epidermal appendages but is restrictive for neural tissue. These results are important for understanding limb development and regenerative medicine strategies.<\/jats:p>","DOI":"10.1111\/ahe.12835","type":"journal-article","created":{"date-parts":[[2022,7,11]],"date-time":"2022-07-11T09:38:02Z","timestamp":1657532282000},"page":"592-601","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Transmembranous and enchondral osteogenesis in transplants of rat limb buds cultivated in serum\u2010 and protein\u2010free culture medium"],"prefix":"10.1111","volume":"51","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2143-6968","authenticated-orcid":false,"given":"Marta","family":"Himelreich Peri\u0107","sequence":"first","affiliation":[{"name":"Center of Excellence in Reproductive and Regenerative Medicine School of Medicine  Zagreb Croatia"},{"name":"Department of Biology School of Medicine  Zagreb Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vedrana","family":"Mu\u017ei\u0107\u2010Radovi\u0107","sequence":"additional","affiliation":[{"name":"Center of Excellence in Reproductive and Regenerative Medicine School of Medicine  Zagreb Croatia"},{"name":"Hospital for Medical Rehabilitation of the Heart and Lung Diseases and Rheumatism \u2010Thalassotherapia Opatija  Opatija Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6238-9811","authenticated-orcid":false,"given":"Tihana","family":"Mari\u0107","sequence":"additional","affiliation":[{"name":"Center of Excellence in Reproductive and Regenerative Medicine School of Medicine  Zagreb Croatia"},{"name":"Department of Biology School of Medicine  Zagreb Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Floriana","family":"Buli\u0107\u2010Jaku\u0161","sequence":"additional","affiliation":[{"name":"Center of Excellence in Reproductive and Regenerative Medicine School of Medicine  Zagreb Croatia"},{"name":"Department of Biology School of Medicine  Zagreb Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gordana","family":"Juri\u0107\u2010Leki\u0107","sequence":"additional","affiliation":[{"name":"Center of Excellence in Reproductive and Regenerative Medicine School of Medicine  Zagreb Croatia"},{"name":"Department of Histology and Embryology School of Medicine  Zagreb Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marta","family":"Takahashi","sequence":"additional","affiliation":[{"name":"Department of Histology and Embryology School of Medicine  Zagreb Croatia"},{"name":"Department of Communicology Catholic University of Croatia  Zagreb Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nino","family":"Sin\u010di\u0107","sequence":"additional","affiliation":[{"name":"Center of Excellence in Reproductive and Regenerative Medicine School of Medicine  Zagreb Croatia"},{"name":"Department of Biology School of Medicine  Zagreb Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Davor","family":"Je\u017eek","sequence":"additional","affiliation":[{"name":"Center of Excellence in Reproductive and Regenerative Medicine School of Medicine  Zagreb Croatia"},{"name":"Department of Histology and Embryology School of Medicine  Zagreb Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana","family":"Katu\u0161i\u0107\u2010Bojanac","sequence":"additional","affiliation":[{"name":"Center of Excellence in Reproductive and Regenerative Medicine School of Medicine  Zagreb Croatia"},{"name":"Department of Biology School of Medicine  Zagreb Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2022,7,11]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.3325\/cmj.2013.54.489"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/wdev.219"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1159\/000047871"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.3109\/10715768909087937"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/boneres.2016.44"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.compositesb.2021.109270"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00204\u2010020\u201002906\u2010z"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/term.2498"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1177\/20417314211004211"},{"volume-title":"Induction and competence. 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