{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,7]],"date-time":"2026-11-07T01:14:53Z","timestamp":1794014093953,"version":"4.8.0"},"reference-count":36,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2020,3,19]],"date-time":"2020-03-19T00:00:00Z","timestamp":1584576000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2020,3,19]],"date-time":"2020-03-19T00:00:00Z","timestamp":1584576000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Aberrant patterns of DNA methylation are consistent events in SCNT derived embryos and mechanistically are believed to be related to abnormal development. While some epigenetic drugs have been used in attempts to improve SCNT efficiency but some concerns remained toward the safety of these drugs on the health of future offspring. Folate is an essential cofactor in one\u2010carbon cycle for conversion of homocysteine to methionine, thereby ensuring supply of SAM, the universal methyl donor for many biological methylation reactions including DNA methylation. Therefore,\n                    <jats:italic>in vitro<\/jats:italic>\n                    DNA hypo-methylation can be induced by folate deprivation and this study aims at deciphering the role of folic acid deprivation in culture medium of BFFs for 6 days on SCNT efficiency. Our data revealed that culture of fibroblast cells in folate\u2212 medium containing 0.5% FBS did not alter the cell cycle compared to other groups. Flowcytometric analysis revealed that DNA methylation (5-mC level) in folate deprived cells cultured in 0.5% serum was decreased compared to folate+ group. The result of bisulfite sequencing was in accordance with flowcytometric analysis, which indicated a decrease in DNA methylation of\n                    <jats:italic>POU5F1<\/jats:italic>\n                    promoter. Gene expression analysis revealed an increase in expression of\n                    <jats:italic>POU5F1<\/jats:italic>\n                    gene in folate\u2212 group. The nuclear area of the cells in folate\u2212 group was significantly larger than folate+ group. Induced DNA hypomethylation by folate deprivation in the folate\u2212 group significantly improved blastocyst rate compared to the folate+ group. DNA methylation level in\n                    <jats:italic>POU5F1<\/jats:italic>\n                    promoter and ICR of\n                    <jats:italic>H19<\/jats:italic>\n                    and\n                    <jats:italic>IGF2<\/jats:italic>\n                    of SCNT derived embryos in the folate\u2212 group was similar to the IVF derived blastocysts. In conclusion, our results proposes a promising \u201cnon-chemical\u201d instead of \u201cchemical\u201d approach using inhibitors of epigenetic modifier enzymes for improving mammalian SCNT efficiency for agricultural and biomedical purposes.\n                  <\/jats:p>","DOI":"10.1038\/s41598-020-61797-3","type":"journal-article","created":{"date-parts":[[2020,3,19]],"date-time":"2020-03-19T07:04:07Z","timestamp":1584601447000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Induced DNA hypomethylation by Folic Acid Deprivation in Bovine Fibroblast Donor Cells Improves Reprogramming of Somatic Cell Nuclear Transfer Embryos"],"prefix":"10.1038","volume":"10","author":[{"given":"Mina","family":"Jozi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Farnoosh","family":"Jafarpour","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reza","family":"Moradi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Faezeh Ghazvini","family":"Zadegan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Khadijeh","family":"Karbalaie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammad Hossein","family":"Nasr-Esfahani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,3,19]]},"reference":[{"key":"61797_CR1","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1111\/asj.12035","volume":"3","author":"S Akagi","year":"2013","unstructured":"Akagi, S., Geshi, M. & Nagai, T. Recent progress in bovine somatic cell nuclear transfer. Anim Sci J. 3, 191\u2013199 (2013).","journal-title":"Anim Sci J."},{"key":"61797_CR2","doi-asserted-by":"publisher","first-page":"810","DOI":"10.1038\/385810a0","volume":"385","author":"I Wilmut","year":"1997","unstructured":"Wilmut, I., Schnieke, A. E., McWhir, J., Kind, A. J. & Campbell, K. H. Viable offspring derived from fetal and adult mammalian cells. Nature 385, 810 (1997).","journal-title":"Nature"},{"key":"61797_CR3","doi-asserted-by":"publisher","first-page":"1869","DOI":"10.1016\/j.theriogenology.2012.05.030","volume":"78","author":"N Rodriguez-Osorio","year":"2012","unstructured":"Rodriguez-Osorio, N., Urrego, R., Cibelli, J., Eilertsen, K. & Memili, E. Reprogramming mammalian somatic cells. Theriogenology 78, 1869\u20131886 (2012).","journal-title":"Theriogenology"},{"key":"61797_CR4","doi-asserted-by":"publisher","first-page":"13734","DOI":"10.1073\/pnas.241522698","volume":"98","author":"W Dean","year":"2001","unstructured":"Dean, W. et al. Conservation of methylation reprogramming in mammalian development: aberrant reprogramming in cloned embryos. Proceedings of the National Academy of Sciences 98, 13734\u201313738 (2001).","journal-title":"Proceedings of the National Academy of Sciences"},{"key":"61797_CR5","doi-asserted-by":"publisher","first-page":"1116","DOI":"10.1016\/S0960-9822(03)00419-6","volume":"13","author":"F Santos","year":"2003","unstructured":"Santos, F. et al. Epigenetic marking correlates with developmental potential in cloned bovine preimplantation embryos. Current Biology 13, 1116\u20131121 (2003).","journal-title":"Current Biology"},{"key":"61797_CR6","doi-asserted-by":"crossref","first-page":"17174","DOI":"10.1016\/S0021-9258(17)44885-X","volume":"265","author":"M Yoshida","year":"1990","unstructured":"Yoshida, M., Kijima, M., Akita, M. & Beppu, T. Potent and specific inhibition of mammalian histone deacetylase both in vivo and in vitro by trichostatin A. Journal of Biological Chemistry 265, 17174\u201317179 (1990).","journal-title":"Journal of Biological Chemistry"},{"key":"61797_CR7","doi-asserted-by":"publisher","first-page":"8674","DOI":"10.1074\/jbc.M009392200","volume":"276","author":"A El Kharroubi","year":"2001","unstructured":"El Kharroubi, A., Piras, G. & Stewart, C. L. DNA demethylation reactivates a subset of imprinted genes in uniparental mouse embryonic fibroblasts. Journal of Biological Chemistry 276, 8674\u20138680 (2001).","journal-title":"Journal of Biological Chemistry"},{"key":"61797_CR8","doi-asserted-by":"publisher","first-page":"896","DOI":"10.1095\/biolreprod.103.017954","volume":"69","author":"BP Enright","year":"2003","unstructured":"Enright, B. P., Kubota, C., Yang, X. & Tian, X. C. Epigenetic characteristics and development of embryos cloned from donor cells treated by trichostatin A or 5-aza-2\u2032-deoxycytidine. Biol Reprod 69, 896\u2013901 (2003).","journal-title":"Biol Reprod"},{"key":"61797_CR9","doi-asserted-by":"publisher","first-page":"2007","DOI":"10.1634\/stemcells.2006-0050","volume":"24","author":"R Blelloch","year":"2006","unstructured":"Blelloch, R. et al. Reprogramming efficiency following somatic cell nuclear transfer is influenced by the differentiation and methylation state of the donor nucleus. Stem Cells 24, 2007\u20132013 (2006).","journal-title":"Stem Cells"},{"key":"61797_CR10","doi-asserted-by":"publisher","first-page":"33094","DOI":"10.1074\/jbc.M112.365478","volume":"287","author":"MB Lee","year":"2012","unstructured":"Lee, M. B. et al. Betaine homocysteine methyltransferase is active in the mouse blastocyst and promotes inner cell mass development. Journal of Biological Chemistry 287, 33094\u201333103 (2012).","journal-title":"Journal of Biological Chemistry"},{"key":"61797_CR11","doi-asserted-by":"publisher","first-page":"184","DOI":"10.1006\/abbi.2001.2474","volume":"393","author":"CV Delgado-Reyes","year":"2001","unstructured":"Delgado-Reyes, C. V., Wallig, M. A. & Garrow, T. A. Immunohistochemical detection of betaine\u2013homocysteine S-methyltransferase in human, pig, and rat liver and kidney. Archives of Biochemistry and Biophysics 393, 184\u2013186 (2001).","journal-title":"Archives of Biochemistry and Biophysics"},{"key":"61797_CR12","doi-asserted-by":"publisher","first-page":"217","DOI":"10.1146\/annurev.nutr.19.1.217","volume":"19","author":"J Selhub","year":"1999","unstructured":"Selhub, J. Homocysteine metabolism. Annual review of nutrition 19, 217\u2013246 (1999).","journal-title":"Annual review of nutrition"},{"key":"61797_CR13","doi-asserted-by":"publisher","first-page":"572","DOI":"10.1038\/nrc3557","volume":"13","author":"JW Locasale","year":"2013","unstructured":"Locasale, J. W. Serine, glycine and one-carbon units: cancer metabolism in full circle. Nature reviews Cancer 13, 572 (2013).","journal-title":"Nature reviews Cancer"},{"key":"61797_CR14","doi-asserted-by":"publisher","first-page":"922","DOI":"10.3390\/nu7020922","volume":"7","author":"KS Bishop","year":"2015","unstructured":"Bishop, K. S. & Ferguson, L. R. The interaction between epigenetics, nutrition and the development of cancer. Nutrients 7, 922\u2013947 (2015).","journal-title":"Nutrients"},{"key":"61797_CR15","doi-asserted-by":"publisher","first-page":"981","DOI":"10.1093\/carcin\/bgi037","volume":"26","author":"JM Stempak","year":"2005","unstructured":"Stempak, J. M., Sohn, K. J., Chiang, E. P., Shane, B. & Kim, Y. I. Cell and stage of transformation-specific effects of folate deficiency on methionine cycle intermediates and DNA methylation in an in vitro model. Carcinogenesis 26, 981\u2013990 (2005).","journal-title":"Carcinogenesis"},{"key":"61797_CR16","doi-asserted-by":"publisher","first-page":"239","DOI":"10.1016\/0009-8981(91)90235-5","volume":"204","author":"J Wang","year":"1991","unstructured":"Wang, J., Dudman, N. P., Lynch, J. & Wilcken, D. E. Betaine: homocysteine methyltransferase\u2014a new assay for the liver enzyme and its absence from human skin fibroblasts and peripheral blood lymphocytes. Clinica Chimica Acta. 204, 239\u2013249 (1991).","journal-title":"Clinica Chimica Acta."},{"key":"61797_CR17","first-page":"64480M","volume":"6448","author":"JL Nadeau","year":"2007","unstructured":"Nadeau, J. L. et al. Quantum dot systems for specific biosensing applications. In Colloidal Quantum Dots for Biomedical Applications II. International Society for Optics and Photonics 6448, 64480M (2007).","journal-title":"International Society for Optics and Photonics"},{"key":"61797_CR18","doi-asserted-by":"publisher","first-page":"371","DOI":"10.1007\/s10620-015-3907-2","volume":"61","author":"D Garc\u00eda-Compe\u00e1n","year":"2016","unstructured":"Garc\u00eda-Compe\u00e1n, D. et al. Current concepts in diabetes mellitus and chronic liver disease: clinical outcomes, hepatitis C virus association, and therapy. Digestive diseases and sciences 61, 371\u2013380 (2016).","journal-title":"Digestive diseases and sciences"},{"key":"61797_CR19","doi-asserted-by":"publisher","first-page":"702","DOI":"10.1016\/j.bbrc.2014.04.082","volume":"447","author":"B Zhu","year":"2014","unstructured":"Zhu, B. et al. MTHFR promotes heterochromatin maintenance. Biochemical and biophysical research communications 447, 702\u2013706 (2014).","journal-title":"Biochemical and biophysical research communications"},{"key":"61797_CR20","doi-asserted-by":"publisher","first-page":"12834","DOI":"10.1073\/pnas.0914328107","volume":"107","author":"MG Mehedint","year":"2010","unstructured":"Mehedint, M. G., Craciunescu, C. N. & Zeisel, S. H. Maternal dietary choline deficiency alters angiogenesis in fetal mouse hippocampus. Proceedings of the National Academy of Sciences 107, 12834\u201312839 (2010).","journal-title":"Proceedings of the National Academy of Sciences"},{"key":"61797_CR21","doi-asserted-by":"publisher","first-page":"3123","DOI":"10.1093\/hmg\/ddg338","volume":"12","author":"F Cerrato","year":"2003","unstructured":"Cerrato, F. et al. Paternal imprints can be established on the maternal Igf2-H19 locus without altering replication timing of DNA. Human molecular genetics 12, 3123\u20133132 (2003).","journal-title":"Human molecular genetics"},{"key":"61797_CR22","doi-asserted-by":"publisher","first-page":"811","DOI":"10.1038\/ng.864","volume":"43","author":"SA Smallwood","year":"2011","unstructured":"Smallwood, S. A. et al. Dynamic CpG island methylation landscape in oocytes and preimplantation embryos. Nature genetics 43, 811 (2011).","journal-title":"Nature genetics"},{"key":"61797_CR23","doi-asserted-by":"publisher","first-page":"5285","DOI":"10.1074\/jbc.M108586200","volume":"277","author":"Y Obata","year":"2002","unstructured":"Obata, Y. & Kono, T. Maternal primary imprinting is established at a specific time for each gene throughout oocyte growth. Journal of Biological Chemistry 277, 5285\u20135289 (2002).","journal-title":"Journal of Biological Chemistry"},{"key":"61797_CR24","doi-asserted-by":"publisher","first-page":"1055","DOI":"10.1097\/00001756-200507130-00005","volume":"16","author":"II Kruman","year":"2005","unstructured":"Kruman, I. I., Mouton, P. R., Emokpae, R. Jr., Cutler, R. G. & Mattson, M. P. Folate deficiency inhibits proliferation of adult hippocampal progenitors. Neuroreport 16, 1055\u20131059 (2005).","journal-title":"Neuroreport"},{"key":"61797_CR25","doi-asserted-by":"publisher","first-page":"162","DOI":"10.1093\/jn\/134.1.162","volume":"134","author":"CN Craciunescu","year":"2004","unstructured":"Craciunescu, C. N. et al. Folic acid deficiency during late gestation decreases progenitor cell proliferation and increases apoptosis in fetal mouse brain. The Journal of nutrition 134, 162\u2013166 (2004).","journal-title":"The Journal of nutrition"},{"key":"61797_CR26","doi-asserted-by":"publisher","first-page":"3186","DOI":"10.4049\/jimmunol.173.5.3186","volume":"173","author":"C Courtemanche","year":"2004","unstructured":"Courtemanche, C., Elson-Schwab, I., Mashiyama, S. T., Kerry, N. & Ames, B. N. Folate deficiency inhibits the proliferation of primary human CD8+ T lymphocytes in vitro. The Journal of Immunology 173, 3186\u20133192 (2004).","journal-title":"The Journal of Immunology"},{"key":"61797_CR27","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1089\/cell.2011.0005","volume":"13","author":"F Jafarpour","year":"2011","unstructured":"Jafarpour, F. et al. Somatic cell-induced hyperacetylation, but not hypomethylation, positively and reversibly affects the efficiency of in vitro cloned blastocyst production in cattle. Cellular Reprogramming (Formerly \u201cCloning and Stem Cells\u201d) 13, 483\u2013493 (2011).","journal-title":"Cellular Reprogramming (Formerly \u201cCloning and Stem Cells\u201d)"},{"key":"61797_CR28","doi-asserted-by":"publisher","first-page":"1013","DOI":"10.1095\/biolreprod60.4.1013","volume":"60","author":"AC Boquest","year":"1999","unstructured":"Boquest, A. C., Day, B. N. & Prather, R. S. Flow cytometric cell cycle analysis of cultured porcine fetal fibroblast cells. Biology of reproduction 60, 1013\u20131019 (1999).","journal-title":"Biology of reproduction"},{"key":"61797_CR29","doi-asserted-by":"publisher","DOI":"10.1186\/s12861-017-0152-2","volume":"17","author":"M Pezhman","year":"2017","unstructured":"Pezhman, M. et al. Cathepsin B inhibitor improves developmental competency and cryo-tolerance of in vitro ovine embryos. BMC developmental biology 17, 10 (2017).","journal-title":"BMC developmental biology"},{"key":"61797_CR30","doi-asserted-by":"publisher","first-page":"168","DOI":"10.1016\/j.cryobiol.2016.08.003","volume":"73","author":"M Derakhshan-Horeh","year":"2016","unstructured":"Derakhshan-Horeh, M. et al. Vitrification at Day3 stage appears not to affect the methylation status of H19\/IGF2 differentially methylated region of in vitro produced human blastocysts. Cryobiology 73, 168\u2013174 (2016).","journal-title":"Cryobiology"},{"key":"61797_CR31","doi-asserted-by":"publisher","first-page":"9821","DOI":"10.1073\/pnas.93.18.9821","volume":"93","author":"JG Herman","year":"1996","unstructured":"Herman, J. G., Graff, J. R., My\u00f6h\u00e4nen, S., Nelkin, B. D. & Baylin, S. B. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proceedings of the national academy of sciences 93, 9821\u20139826 (1996).","journal-title":"Proceedings of the national academy of sciences"},{"key":"61797_CR32","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-11-230","volume":"11","author":"C Rohde","year":"2010","unstructured":"Rohde, C., Zhang, Y., Reinhardt, R. & Jeltsch, A. BISMA-Fast and accurate bisulfite sequencing data analysis of individual clones from unique and repetitive sequences. BMC bioinformatics 11, 230 (2010).","journal-title":"BMC bioinformatics"},{"key":"61797_CR33","doi-asserted-by":"publisher","DOI":"10.1186\/s12861-017-0152-2","volume":"17","author":"M Pezhman","year":"2017","unstructured":"Pezhman, M. et al. Cathepsin B inhibitor improves developmental competency and cryo-tolerance of in vitro ovine embryos. BMC developmental biology 17, 10 (2017).","journal-title":"BMC developmental biology"},{"key":"61797_CR34","doi-asserted-by":"publisher","first-page":"34","DOI":"10.1016\/j.cryobiol.2017.06.001","volume":"77","author":"F Sefid","year":"2017","unstructured":"Sefid, F. et al. Vitamin K2 improves developmental competency and cryo-tolerance of in vitro derived ovine blastocyst. Cryobiology 77, 34\u201340 (2017).","journal-title":"Cryobiology"},{"key":"61797_CR35","doi-asserted-by":"publisher","DOI":"10.1186\/s12864-015-2264-z","volume":"17","author":"SM Hosseini","year":"2016","unstructured":"Hosseini, S. M. et al. Epigenetic modification with trichostatin A does not correct specific errors of somatic cell nuclear transfer at the transcriptomic level; highlighting the non-random nature of oocyte-mediated reprogramming errors. BMC genomics 17, 16 (2016).","journal-title":"BMC genomics"},{"key":"61797_CR36","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1089\/cell.2012.0033","volume":"15","author":"SM Hosseini","year":"2013","unstructured":"Hosseini, S. M. et al. Cloned sheep blastocysts derived from oocytes enucleated manually using a pulled pasteur pipette. Cellular Reprogramming (Formerly \u201cCloning and Stem Cells\u201d) 15, 15\u201323 (2013).","journal-title":"Cellular Reprogramming (Formerly \u201cCloning and Stem Cells\u201d)"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-61797-3.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-61797-3","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-61797-3.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,6]],"date-time":"2022-12-06T03:22:36Z","timestamp":1670296956000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-61797-3"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,3,19]]},"references-count":36,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2020,12]]}},"alternative-id":["61797"],"URL":"https:\/\/doi.org\/10.1038\/s41598-020-61797-3","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,3,19]]},"assertion":[{"value":"21 June 2019","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"2 March 2020","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 March 2020","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"5076"}}