{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T19:47:52Z","timestamp":1787687272246,"version":"build-2784847793"},"reference-count":34,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2024,9,12]],"date-time":"2024-09-12T00:00:00Z","timestamp":1726099200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100007049","name":"Korea Institute of Ocean Science and Technology","doi-asserted-by":"publisher","award":["PEA0215"],"award-info":[{"award-number":["PEA0215"]}],"id":[{"id":"10.13039\/501100007049","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The development of serum-free media (SFM) is critical to advance cell culture techniques used in viral vaccine production and address the ethical concerns and contamination risks associated with fetal bovine serum (FBS). This study evaluated the effects of marine microalgal extracts and growth factor cocktails on the activity of Madin-Darby canine kidney (MDCK) and Vero cells. Five marine microalgal species were used: Spirulina platensis (SP), Dunaliella salina (DS), Haematococcus pluvialis (HP), Nannochloropsis salina (NS), and Tetraselmis sp. (TS). DS and SP extracts significantly increased the proliferation rate of both MDCK and Vero cells. DS had a proliferation rate of 149.56% and 195.50% in MDCK and Vero cells, respectively, compared with that in serum-free medium (SFM). Notably, DS and SP extracts significantly increased superoxide dismutase (SOD) activity, which was 118.61% in MDCK cells and 130.08% in Vero cells for DS, and 108.72% in MDCK cells and 125.63% in Vero cells for SP, indicating a reduction in intracellular oxidative stress. Marine microalgal extracts, especially DS and SP, are feasible alternatives to FBS in cell culture as they promote cell proliferation, ensure safety, and supply essential nutrients while reducing oxidative stress.<\/jats:p>","DOI":"10.3390\/ijms25189881","type":"journal-article","created":{"date-parts":[[2024,9,13]],"date-time":"2024-09-13T03:03:37Z","timestamp":1726196617000},"page":"9881","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Serum-Free Media Formulation Using Marine Microalgae Extracts and Growth Factor Cocktails for Madin-Darby Canine Kidney and Vero Cell Cultures"],"prefix":"10.3390","volume":"25","author":[{"given":"Areumi","family":"Park","sequence":"first","affiliation":[{"name":"Jeju Bio Research Center, Korea Institute of Ocean Science and Technology (KIOST), Jeju 63349, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7772-0207","authenticated-orcid":false,"given":"Yeon-Ji","family":"Lee","sequence":"additional","affiliation":[{"name":"Jeju Bio Research Center, Korea Institute of Ocean Science and Technology (KIOST), Jeju 63349, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eunyoung","family":"Jo","sequence":"additional","affiliation":[{"name":"Jeju Bio Research Center, Korea Institute of Ocean Science and Technology (KIOST), Jeju 63349, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7860-3459","authenticated-orcid":false,"given":"Gun-Hoo","family":"Park","sequence":"additional","affiliation":[{"name":"Jeju Bio Research Center, Korea Institute of Ocean Science and Technology (KIOST), Jeju 63349, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seong-Yeong","family":"Heo","sequence":"additional","affiliation":[{"name":"Jeju Bio Research Center, Korea Institute of Ocean Science and Technology (KIOST), Jeju 63349, Republic of Korea"},{"name":"Department of Marine Technology & Convergence Engineering (Marine Biotechnology), University of Science and Technology (UST), Daejeon 34113, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-6402-7712","authenticated-orcid":false,"given":"Eun-Jeong","family":"Koh","sequence":"additional","affiliation":[{"name":"Jeju Bio Research Center, Korea Institute of Ocean Science and Technology (KIOST), Jeju 63349, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seung-Hong","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Engineering, Soonchunhyang University, Asan 31538, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5004-3223","authenticated-orcid":false,"given":"Seon-Heui","family":"Cha","sequence":"additional","affiliation":[{"name":"Department of Marine Bio and Medical Sciences, Hanseo University, Seosan-si 32158, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soo-Jin","family":"Heo","sequence":"additional","affiliation":[{"name":"Jeju Bio Research Center, Korea Institute of Ocean Science and Technology (KIOST), Jeju 63349, Republic of Korea"},{"name":"Department of Marine Technology & Convergence Engineering (Marine Biotechnology), University of Science and Technology (UST), Daejeon 34113, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,9,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1586\/erv.09.19","article-title":"Vero cell platform in vaccine production: Moving towards cell culture-based viral vaccines","volume":"8","author":"Barrett","year":"2009","journal-title":"Expert. Rev. Vaccines"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"9","DOI":"10.4167\/jbv.2013.43.1.9","article-title":"Cell culture-based influenza vaccines as alternatives to egg-based vaccines","volume":"43","author":"Lee","year":"2013","journal-title":"J. Bacteriol. Virol."},{"key":"ref_3","unstructured":"Grosvenor, S. (2008). The role of media development in process optimization: An historical perspective. Biopharm. Int., 2008."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"998","DOI":"10.1128\/CVI.00024-13","article-title":"An MDCK cell culture-derived formalin-inactivated influenza virus whole-virion vaccine from an influenza virus library confers cross-protective immunity by intranasal administration in mice","volume":"20","author":"Haredy","year":"2013","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1007\/s00253-010-2742-9","article-title":"MDCK and Vero cells for influenza virus vaccine production: A one-to-one comparison up to lab-scale bioreactor cultivation","volume":"88","author":"Genzel","year":"2010","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_6","first-page":"16904","article-title":"Animal cell lines as expression platforms in viral vaccine production: A post COVID-19 perspective","volume":"9","author":"Demirden","year":"2024","journal-title":"ACS Omega"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1080\/14760584.2023.2214219","article-title":"Suspended cell lines for inactivated virus vaccine production","volume":"22","author":"Zhang","year":"2023","journal-title":"Expert. Rev. Vaccines"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Yamanaka, K., Haraguchi, Y., Takahashi, H., Kawashima, I., and Shimizu, T. (2023). Development of serum-free and grain-derived-nutrient-free medium using microalga-derived nutrients and mammalian cell-secreted growth factors for sustainable cultured meat production. Sci. Rep., 13.","DOI":"10.1038\/s41598-023-27629-w"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"6064","DOI":"10.1021\/acs.jafc.4c00624","article-title":"Integrated omics approach: Revealing the mechanism of Auxenochlorella pyrenoidosa protein extract replacing fetal bovine serum for fish muscle cell culture","volume":"72","author":"Dong","year":"2024","journal-title":"J. Agric. Food Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.jcyt.2016.11.011","article-title":"A consensus introduction to serum replacements and serum-free media for cellular therapies","volume":"19","author":"Karnieli","year":"2017","journal-title":"Cytotherapy"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"48","DOI":"10.7774\/cevr.2020.9.1.48","article-title":"Suspension culture of Vero cells for the production of adenovirus type 5","volume":"9","author":"Lee","year":"2020","journal-title":"Clin. Exp. Vaccine Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.jviromet.2010.09.029","article-title":"Adaptation of a Madin\u2013Darby canine kidney cell line to suspension growth in serum-free media and comparison of its ability to produce avian influenza virus to Vero and BHK21 cell lines","volume":"171","author":"Harmsen","year":"2011","journal-title":"J. Virol. Methods"},{"key":"ref_13","unstructured":"Lee, H.C. (1995). Serum-Free Medium in Animal Cell Culture, BT NEWS."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Kolkmann, A.M., Van Essen, A., Post, M.J., and Moutsatsou, P. (2022). Development of a chemically defined medium for in vitro expansion of primary bovine satellite cells. Front. Bioeng. Biotechnol., 10.","DOI":"10.3389\/fbioe.2022.895289"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Li, K., Zhang, X., Wang, D., Tuan, R.S., and Ker, D.F.E. (2023). Synergistic effects of growth factor-based serum-free medium and tendon-like substrate topography on tenogenesis of mesenchymal stem cells. Biomater. Adv., 146.","DOI":"10.1016\/j.bioadv.2023.213316"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1263\/jbb.101.87","article-title":"Commercial applications of microalgae","volume":"101","author":"Spolaore","year":"2006","journal-title":"J. Biosci. Bioeng."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Okamoto, Y., Haraguchi, Y., Sawamura, N., Asahi, T., and Shimizu, T. (2020). Mammalian cell cultivation using nutrients extracted from microalgae. Biotechnol. Prog., 36.","DOI":"10.1002\/btpr.2941"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Jeong, Y., Choi, W.Y., Park, A., Lee, Y.J., Lee, Y., Park, G.H., Lee, S.J., Lee, W.K., Ryu, Y.K., and Kang, D.H. (2021). Marine cyanobacterium Spirulina maxima as an alternate to the animal cell culture medium supplement. Sci. Rep., 11.","DOI":"10.1038\/s41598-021-84558-2"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Defendi-Cho, G., and Gould, T.M. (2023). In vitro culture of bovine fibroblasts using select serum-free media supplemented with Chlorella vulgaris extract. BMC Biotechnol., 23.","DOI":"10.1186\/s12896-023-00774-w"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"636","DOI":"10.3892\/ijmm.2012.1028","article-title":"Wnt3a inhibits proliferation but promotes melanogenesis of melan-a cells","volume":"30","author":"Guo","year":"2012","journal-title":"Int. J. Mol. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"415","DOI":"10.4163\/jnh.2017.50.5.415","article-title":"Neuroprotective effects of Momordica charantia extract against hydrogen peroxide-induced cytotoxicity in human neuroblastoma SK-N-MC cells","volume":"50","author":"Kim","year":"2017","journal-title":"J. Nutr. Health"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1161\/01.RES.85.2.147","article-title":"Inhibition of copper-zinc superoxide dismutase induces cell growth, hypertrophic phenotype, and apoptosis in neonatal rat cardiac myocytes in vitro","volume":"85","author":"Siwik","year":"1999","journal-title":"Circ. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.fshw.2019.03.001","article-title":"Microalgae: A potential alternative to health supplementation for humans","volume":"8","author":"Koyande","year":"2019","journal-title":"Food Sci. Hum. Wellness"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1016\/S0021-9258(18)65826-0","article-title":"The growth response of mammalian cells in tissue culture to L-glutamine and L-glutamic acid","volume":"218","author":"Eagle","year":"1956","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Wang, X., Zhang, C., and Bao, N. (2023). Molecular mechanism of palmitic acid and its derivatives in tumor progression. Front. Oncol., 13.","DOI":"10.3389\/fonc.2023.1224125"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Conte, F., van Buuringen, N., Voermans, N.C., and Lefeber, D.J. (2021). Galactose in human metabolism, glycosylation and congenital metabolic diseases: Time for a closer look. Biochim. Biophys. Acta Gen. Subj., 1865.","DOI":"10.1016\/j.bbagen.2021.129898"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"102455","DOI":"10.1016\/j.canep.2023.102455","article-title":"Association of serum superoxide dismutase activity and the incidence of colorectal cancer in a nested case-control study","volume":"87","author":"Adachi","year":"2023","journal-title":"Cancer Epidemiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2032","DOI":"10.12659\/MSM.913375","article-title":"Evaluation and monitoring of superoxide dismutase (SOD) activity and its clinical significance in gastric cancer: A systematic review and meta-analysis","volume":"25","author":"Li","year":"2019","journal-title":"Med. Sci. Monit."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1016\/j.freeradbiomed.2008.05.023","article-title":"Prooxidant activity of the superoxide dismutase (SOD)-mimetic EUK-8 in proliferating and growth-arrested Escherichia coli cells","volume":"45","author":"Matthijssens","year":"2008","journal-title":"Free Radic. Biol. Med."},{"key":"ref_30","first-page":"1","article-title":"Rapid, accurate, sensitive, and reproducible HPLC analysis of amino acids","volume":"1100","author":"Henderson","year":"2000","journal-title":"Amino Acid. Anal. Using. Zorbax Eclipse-AAA Columns Agil."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1006\/abio.1993.1244","article-title":"One-step lipid extraction and fatty acid methyl esters preparation from fresh plant tissues","volume":"211","author":"Mancha","year":"1993","journal-title":"Anal. Biochem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"8214","DOI":"10.1039\/D1FO03946F","article-title":"Structural characteristics of sulfated polysaccharides from Sargassum horneri and immune-enhancing activity of polysaccharides combined with lactic acid bacteria","volume":"13","author":"Kim","year":"2022","journal-title":"Food Funct."},{"key":"ref_33","first-page":"281","article-title":"Development of a PCR assay to detect mycoplasma contamination in cord blood hematopoietic stem cells","volume":"6","author":"Fazaely","year":"2014","journal-title":"Iran. J. Microbiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1823","DOI":"10.1016\/j.theriogenology.2005.04.016","article-title":"Comparison of cell proliferation index in equine and caprine embryos using a modified BrdU incorporation assay","volume":"64","author":"Moussa","year":"2005","journal-title":"Theriogenology"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/18\/9881\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:55:07Z","timestamp":1760111707000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/18\/9881"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,9,12]]},"references-count":34,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2024,9]]}},"alternative-id":["ijms25189881"],"URL":"https:\/\/doi.org\/10.3390\/ijms25189881","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,9,12]]}}}