{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T09:50:16Z","timestamp":1791798616470,"version":"4.3.4"},"reference-count":51,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,2,1]],"date-time":"2022-02-01T00:00:00Z","timestamp":1643673600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100017501","name":"Science-Health Joint Medical Scientific Research Project of Chongqing","doi-asserted-by":"publisher","award":["2021MSXM072"],"award-info":[{"award-number":["2021MSXM072"]}],"id":[{"id":"10.13039\/100017501","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Genet."],"abstract":"<jats:p>Although vitrification has been widely applied in assisted reproductive technology, it is unknown whether storage time has any impact on the mRNA and lncRNA expression profiles in human embryos. Eleven women (aged 23\u201335\u00a0years) who had undergone <jats:italic>in vitro<\/jats:italic> fertilization treatment were recruited for this study. The transcriptomes of 3 fresh eight-cell embryos and 8 surviving vitrified-warmed eight-cell embryos (4 embryos were cryostored for 3\u00a0years, and the others were cryostored for 8\u00a0years) were analyzed through single-cell RNA-Seq. No differentially expressed mRNAs or lncRNAs were identified between the 3-years group and 8-years group. A total of 128 mRNAs and 365 lncRNAs were differentially expressed in the 8 vitrified-warmed embryos compared with the fresh embryos. The vitrification-warming impact was moderate, and it was mainly related to the pathways of metabolism, stress response, apoptosis, cell cycle, cell adhesion, and signaling for TFG-\u03b2 and Hippo. The analysis of target mRNAs suggested that lncRNAs might contribute to the regulation of mRNAs after vitrification-warming. Our findings indicated that long-term storage after vitrification does not affect the mRNA and lncRNA expression profiles in human embryos, however, the procedure of vitrification-warming would lead to minor alteration of transcriptome.<\/jats:p>","DOI":"10.3389\/fgene.2021.751467","type":"journal-article","created":{"date-parts":[[2022,2,1]],"date-time":"2022-02-01T05:40:01Z","timestamp":1643694001000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":15,"title":["Long-Term Storage Does Not Affect the Expression Profiles of mRNA and Long Non-Coding RNA in Vitrified-Warmed Human Embryos"],"prefix":"10.3389","volume":"12","author":[{"given":"Jingyu","family":"Li","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ling","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weiwei","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Han","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guoning","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,2,1]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1016\/j.rbmo.2009.11.013","article-title":"Slow Freezing, Vitrification and Ultra-rapid Freezing of Human Embryos: a Systematic Review and Meta-Analysis","volume":"20","author":"AbdelHafez","year":"2010","journal-title":"Reprod. BioMedicine Online"},{"key":"B2","doi-asserted-by":"publisher","first-page":"613","DOI":"10.1002\/mrd.22068","article-title":"Gene Expression Profiles of Vitrified In Vitro- and In Vivo-derived Bovine Blastocysts","volume":"79","author":"Aksu","year":"2012","journal-title":"Mol. Reprod. Dev."},{"key":"B3","doi-asserted-by":"publisher","first-page":"4593","DOI":"10.1091\/mbc.e06-05-0377","article-title":"The Mitochondrial Fission Protein hFis1 Requires the Endoplasmic Reticulum Gateway to Induce Apoptosis","volume":"17","author":"Alirol","year":"2006","journal-title":"MBoC"},{"key":"B4","doi-asserted-by":"publisher","first-page":"1222","DOI":"10.3390\/ijms22031222","article-title":"Effects of Vitrification on the Blastocyst Gene Expression Profile in a Porcine Model","volume":"22","author":"Cuello","year":"2021","journal-title":"Ijms"},{"key":"B5","doi-asserted-by":"publisher","first-page":"44","DOI":"10.1038\/ng.3449","article-title":"The Primate-specific Noncoding RNA HPAT5 Regulates Pluripotency during Human Preimplantation Development and Nuclear Reprogramming","volume":"48","author":"Durruthy-Durruthy","year":"2015","journal-title":"Nat. Genet."},{"key":"B6","doi-asserted-by":"publisher","first-page":"536","DOI":"10.1093\/humupd\/dms016","article-title":"A Critical Appraisal of Cryopreservation (Slow Cooling versus Vitrification) of Human Oocytes and Embryos","volume":"18","author":"Edgar","year":"2012","journal-title":"Hum. Reprod. Update"},{"key":"B7","doi-asserted-by":"publisher","first-page":"392","DOI":"10.1002\/mrd.22042","article-title":"The Importance of Mitochondrial Metabolic Activity and Mitochondrial DNA Replication during Oocyte Maturation In Vitro on Oocyte Quality and Subsequent Embryo Developmental Competence","volume":"79","author":"Ge","year":"2012","journal-title":"Mol. Reprod. Dev."},{"key":"B8","doi-asserted-by":"publisher","first-page":"20","DOI":"10.1007\/BF01131376","article-title":"Long-term Storage of Eight-Cell Mouse Embryos at -196 Degree C","volume":"3","author":"Glenister","year":"1986","journal-title":"J. Assist. Reprod. Genet."},{"key":"B9","doi-asserted-by":"publisher","first-page":"244","DOI":"10.1016\/j.fertnstert.2016.06.029","article-title":"Cleavage-stage or Blastocyst Transfer: what Are the Benefits and Harms?","volume":"106","author":"Glujovsky","year":"2016","journal-title":"Fertil. Sterility"},{"key":"B10","doi-asserted-by":"publisher","first-page":"CD002118","DOI":"10.1002\/14651858.CD002118.pub5","article-title":"Cleavage Stage versus Blastocyst Stage Embryo Transfer in Assisted Reproductive Technology","volume":"6","author":"Glujovsky","year":"2016","journal-title":"Cochrane Database Syst. Rev."},{"key":"B11","doi-asserted-by":"publisher","first-page":"264","DOI":"10.1016\/j.fertnstert.2011.06.029","article-title":"Cryopreservation: a Cold Look at Technology for Fertility Preservation","volume":"96","author":"Gosden","year":"2011","journal-title":"Fertil. Sterility"},{"key":"B12","doi-asserted-by":"publisher","first-page":"e0187268","DOI":"10.1371\/journal.pone.0187268","article-title":"Transcriptomic Difference in Bovine Blastocysts Following Vitrification and Slow Freezing at Morula Stage","volume":"12","author":"Gupta","year":"2017","journal-title":"PLoS One"},{"key":"B13","doi-asserted-by":"publisher","first-page":"338","DOI":"10.1071\/RD20285","article-title":"Effect of Vitrification on Global Gene Expression Dynamics of Bovine Elongating Embryos","volume":"33","author":"Gutierrez-Castillo","year":"2021","journal-title":"Reprod. Fertil. Dev."},{"key":"B14","doi-asserted-by":"publisher","first-page":"264","DOI":"10.1038\/s41586-020-3027-9","article-title":"Reconstitution of the Oocyte Transcriptional Network with Transcription Factors","volume":"589","author":"Hamazaki","year":"2020","journal-title":"Nature"},{"key":"B15","doi-asserted-by":"publisher","first-page":"910","DOI":"10.1242\/dev.116996","article-title":"Gene Activation-Associated Long Noncoding RNAs Function in Mouse Preimplantation Development","volume":"142","author":"Hamazaki","year":"2015","journal-title":"Development"},{"key":"B16","doi-asserted-by":"publisher","first-page":"144","DOI":"10.1007\/s11684-020-0792-7","article-title":"Effects of Vitrification and Cryostorage Duration on Single-Cell RNA-Seq Profiling of Vitrified-Thawed Human Metaphase II Oocytes","volume":"15","author":"Huo","year":"2020","journal-title":"Front. Med."},{"key":"B17","doi-asserted-by":"publisher","first-page":"e0179259","DOI":"10.1371\/journal.pone.0179259","article-title":"Dose-dependent Effects of Gamma Radiation on the Early Zebrafish Development and Gene Expression","volume":"12","author":"Hurem","year":"2017","journal-title":"PLoS One"},{"key":"B18","doi-asserted-by":"publisher","first-page":"197","DOI":"10.1016\/j.jdermsci.2012.09.007","article-title":"Multipotential Functions of Hic-5 in Growth, Differentiation, Migration and Adhesion of Human Keratinocytes","volume":"68","author":"Inui","year":"2012","journal-title":"J. Dermatol. Sci."},{"key":"B19","doi-asserted-by":"publisher","first-page":"2136","DOI":"10.1093\/oxfordjournals.humrep.a136248","article-title":"Fertilization and Early Embryology: Cryopreservation: the Problem of Evaluation","volume":"10","author":"Jones","year":"1995","journal-title":"Hum. Reprod."},{"key":"B20","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1093\/humupd\/dmu063","article-title":"The Effect of Cryopreservation on the Genome of Gametes and Embryos: Principles of Cryobiology and Critical Appraisal of the Evidence","volume":"21","author":"Kopeika","year":"2015","journal-title":"Hum. Reprod. Update"},{"key":"B21","doi-asserted-by":"publisher","first-page":"300","DOI":"10.1016\/s1472-6483(10)60837-1","article-title":"Highly Efficient Vitrification Method for Cryopreservation of Human Oocytes","volume":"11","author":"Kuwayama","year":"2005","journal-title":"Reprod. BioMedicine Online"},{"key":"B22","doi-asserted-by":"publisher","first-page":"652","DOI":"10.1016\/j.theriogenology.2011.03.018","article-title":"Does Storage Time in LN2 Influence Survival and Pregnancy Outcome of Vitrified Rabbit Embryos?","volume":"76","author":"Lavara","year":"2011","journal-title":"Theriogenology"},{"key":"B23","doi-asserted-by":"publisher","first-page":"845","DOI":"10.1002\/bies.10137","article-title":"Quiet Please, Do Not Disturb: a Hypothesis of Embryo Metabolism and Viability","volume":"24","author":"Leese","year":"2002","journal-title":"Bioessays"},{"key":"B24","doi-asserted-by":"publisher","first-page":"e9738","DOI":"10.7717\/peerj.9738","article-title":"Comparing Transcriptome Profiles of Human Embryo Cultured in Closed and Standard Incubators","volume":"8","author":"Li","year":"","journal-title":"PeerJ"},{"key":"B25","doi-asserted-by":"publisher","first-page":"1675","DOI":"10.1093\/humrep\/deaa136","article-title":"The Effect of Storage Time after Vitrification on Pregnancy and Neonatal Outcomes Among 24 698 Patients Following the First Embryo Transfer Cycles","volume":"35","author":"Li","year":"","journal-title":"Hum. Reprod."},{"key":"B26","doi-asserted-by":"publisher","first-page":"96","DOI":"10.1080\/09513590.2016.1207166","article-title":"Influence of Storage Time on Vitrified Human Cleavage-Stage Embryos Froze in Open System","volume":"33","author":"Li","year":"2017","journal-title":"Gynecol. Endocrinol."},{"key":"B27","doi-asserted-by":"publisher","first-page":"471","DOI":"10.1007\/s10815-014-0197-0","article-title":"The Safety of Long-Term Cryopreservation on Slow-Frozen Early Cleavage Human Embryos","volume":"31","author":"Liu","year":"2014","journal-title":"J. Assist. Reprod. Genet."},{"key":"B28","doi-asserted-by":"publisher","first-page":"1367","DOI":"10.1007\/s10815-013-0062-6","article-title":"Embryo Developmental Capability and Pregnancy Outcome Are Related to the Mitochondrial DNA Copy Number and Ooplasmic Volume","volume":"30","author":"Murakoshi","year":"2013","journal-title":"J. Assist. Reprod. Genet."},{"key":"B29","doi-asserted-by":"publisher","first-page":"2681","DOI":"10.1093\/humrep\/dei153","article-title":"Number of Embryos for Transfer after IVF and ICSI: a Cochrane Review","volume":"20","author":"Pandian","year":"2005","journal-title":"Hum. Reprod."},{"key":"B30","doi-asserted-by":"publisher","first-page":"528","DOI":"10.1038\/s41586-019-1469-8","article-title":"Molecular Architecture of Lineage Allocation and Tissue Organization in Early Mouse Embryo","volume":"572","author":"Peng","year":"2019","journal-title":"Nature"},{"key":"B31","doi-asserted-by":"publisher","first-page":"139","DOI":"10.1093\/humupd\/dmw038","article-title":"Oocyte, Embryo and Blastocyst Cryopreservation in ART: Systematic Review and Meta-Analysis Comparing Slow-Freezing versus Vitrification to Produce Evidence for the Development of Global Guidance","volume":"23","author":"Rienzi","year":"2017","journal-title":"Hum. Reprod. Update"},{"key":"B32","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1016\/j.fertnstert.2008.09.084","article-title":"Does Storage Time Influence Postthaw Survival and Pregnancy Outcome? an Analysis of 11,768 Cryopreserved Human Embryos","volume":"93","author":"Riggs","year":"2010","journal-title":"Fertil. Sterility"},{"key":"B33","doi-asserted-by":"publisher","first-page":"7294","DOI":"10.26355\/eurrev_202007_21884","article-title":"TGF\u03b21I1 Suppressed Cell Migration and Invasion in Colorectal Cancer by Inhibiting the TGF-\u03b2 Pathway and EMT Progress","volume":"24","author":"Ruan","year":"2020","journal-title":"Eur. Rev. Med. Pharmacol. Sci."},{"key":"B34","doi-asserted-by":"publisher","first-page":"569","DOI":"10.1530\/REP-12-0047","article-title":"Comparison of Gene Expression in Fresh and Frozen-Thawed Human Preimplantation Embryos","volume":"144","author":"Shaw","year":"2012","journal-title":"Reproduction"},{"key":"B35","doi-asserted-by":"publisher","first-page":"1211","DOI":"10.1016\/j.theriogenology.2010.11.033","article-title":"Lipid Content and Apoptosis of In Vitro-produced Bovine Embryos as Determinants of Susceptibility to Vitrification","volume":"75","author":"Sudano","year":"2011","journal-title":"Theriogenology"},{"key":"B36","doi-asserted-by":"publisher","first-page":"707","DOI":"10.1038\/305707a0","article-title":"Human Pregnancy Following Cryopreservation, Thawing and Transfer of an Eight-Cell Embryo","volume":"305","author":"Trounson","year":"1983","journal-title":"Nature"},{"key":"B37","doi-asserted-by":"publisher","first-page":"614","DOI":"10.1016\/j.rbmo.2018.03.008","article-title":"Cryostorage Duration Does Not Affect Pregnancy and Neonatal Outcomes: a Retrospective Single-centre Cohort Study of Vitrified-Warmed Blastocysts","volume":"36","author":"Ueno","year":"2018","journal-title":"Reprod. BioMedicine Online"},{"key":"B38","doi-asserted-by":"publisher","first-page":"415","DOI":"10.1093\/oxfordjournals.humrep.a135954","article-title":"Fertilization and Early Embryolgoy: ATP Content of Human Oocytes and Developmental Potential and Outcome after In-Vitro Fertilization and Embryo Transfer","volume":"10","author":"Van Blerkom","year":"1995","journal-title":"Hum. Reprod."},{"key":"B39","doi-asserted-by":"publisher","first-page":"1452","DOI":"10.15252\/embr.201642051","article-title":"A Novel Long Intergenic Noncoding RNA Indispensable for the Cleavage of Mouse Two\u2010cell Embryos","volume":"17","author":"Wang","year":"2016","journal-title":"EMBO Rep."},{"key":"B40","doi-asserted-by":"publisher","first-page":"1887","DOI":"10.1016\/j.cell.2018.11.039","article-title":"Asymmetric Expression of LincGET Biases Cell Fate in Two-Cell Mouse Embryos","volume":"175","author":"Wang","year":"2018","journal-title":"Cell"},{"key":"B41","doi-asserted-by":"publisher","first-page":"2729","DOI":"10.1016\/j.fertnstert.2010.03.025","article-title":"Embryo Vitrification Affects the Methylation of the H19\/Igf2 Differentially Methylated Domain and the Expression of H19 and Igf2","volume":"93","author":"Wang","year":"2010","journal-title":"Fertil. Sterility"},{"key":"B42","doi-asserted-by":"publisher","first-page":"1494","DOI":"10.1101\/gad.1800909","article-title":"Long Noncoding RNAs: Functional Surprises from the RNA World","volume":"23","author":"Wilusz","year":"2009","journal-title":"Genes Dev."},{"key":"B43","doi-asserted-by":"publisher","first-page":"2950","DOI":"10.1093\/humrep\/det361","article-title":"The Time Aspect in Storing Vitrified Blastocysts: its Impact on Survival Rate, Implantation Potential and Babies Born","volume":"28","author":"Wirleitner","year":"2013","journal-title":"Hum. Reprod."},{"key":"B44","doi-asserted-by":"publisher","first-page":"329","DOI":"10.1016\/j.devcel.2009.02.012","article-title":"TGF-\u03b2 Superfamily Signaling in Embryonic Development and Homeostasis","volume":"16","author":"Wu","year":"2009","journal-title":"Develop. Cel"},{"key":"B45","doi-asserted-by":"publisher","first-page":"114","DOI":"10.1080\/14647273.2016.1186848","article-title":"Shortened Equilibration Time Can Compromise Clinical Outcomes in Human Embryo Vitrification","volume":"19","author":"Xiong","year":"2016","journal-title":"Hum. Fertil."},{"key":"B46","doi-asserted-by":"publisher","first-page":"593","DOI":"10.1038\/nature12364","article-title":"Genetic Programs in Human and Mouse Early Embryos Revealed by Single-Cell RNA Sequencing","volume":"500","author":"Xue","year":"2013","journal-title":"Nature"},{"key":"B47","doi-asserted-by":"publisher","first-page":"1131","DOI":"10.1038\/nsmb.2660","article-title":"Single-cell RNA-Seq Profiling of Human Preimplantation Embryos and Embryonic Stem Cells","volume":"20","author":"Yan","year":"2013","journal-title":"Nat. Struct. Mol. Biol."},{"key":"B48","doi-asserted-by":"publisher","first-page":"811","DOI":"10.1016\/j.cell.2015.10.044","article-title":"Hippo Pathway in Organ Size Control, Tissue Homeostasis, and Cancer","volume":"163","author":"Yu","year":"2015","journal-title":"Cell"},{"key":"B49","doi-asserted-by":"publisher","first-page":"256","DOI":"10.1016\/j.fertnstert.2013.03.009","article-title":"Effect of Vitrification on Promoter CpG Island Methylation Patterns and Expression Levels of DNA Methyltransferase 1o, Histone Acetyltransferase 1, and Deacetylase 1 in Metaphase II Mouse Oocytes","volume":"100","author":"Zhao","year":"2013","journal-title":"Fertil. Sterility"},{"key":"B50","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1016\/j.cryobiol.2016.08.011","article-title":"Vitrification Transiently Alters Oct-4, Bcl2 and P53 Expression in Mouse Morulae but Does Not Affect Embryo Development In Vitro","volume":"73","author":"Zhou","year":"2016","journal-title":"Cryobiology"},{"key":"B51","doi-asserted-by":"publisher","first-page":"924","DOI":"10.1093\/humrep\/dey044","article-title":"Live Birth Rates in the First Complete IVF Cycle Among 20 687 Women Using a Freeze-All Strategy","volume":"33","author":"Zhu","year":"2018","journal-title":"Hum. Reprod."}],"container-title":["Frontiers in Genetics"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fgene.2021.751467\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,1]],"date-time":"2022-02-01T05:40:06Z","timestamp":1643694006000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fgene.2021.751467\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,1]]},"references-count":51,"alternative-id":["10.3389\/fgene.2021.751467"],"URL":"https:\/\/doi.org\/10.3389\/fgene.2021.751467","relation":{},"ISSN":["1664-8021"],"issn-type":[{"value":"1664-8021","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,1]]},"article-number":"751467"}}