{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,12]],"date-time":"2026-02-12T11:58:51Z","timestamp":1770897531502,"version":"3.50.1"},"reference-count":47,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2024,12,23]],"date-time":"2024-12-23T00:00:00Z","timestamp":1734912000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"China Scholarship Council","award":["202308230130"],"award-info":[{"award-number":["202308230130"]}]},{"name":"China Scholarship Council","award":["202208080057"],"award-info":[{"award-number":["202208080057"]}]},{"name":"China Scholarship Council","award":["KP-06-H81\/9"],"award-info":[{"award-number":["KP-06-H81\/9"]}]},{"name":"Bulgarian National Science Fund","award":["202308230130"],"award-info":[{"award-number":["202308230130"]}]},{"name":"Bulgarian National Science Fund","award":["202208080057"],"award-info":[{"award-number":["202208080057"]}]},{"name":"Bulgarian National Science Fund","award":["KP-06-H81\/9"],"award-info":[{"award-number":["KP-06-H81\/9"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Ovarian tissue cryopreservation has been gradually applied. It is essential to elucidate the differences between cryopreserved and fresh ovarian tissue and to refine cryopreservation protocols for improved outcomes. To explore the transcriptomic differences between fresh ovarian tissue and tissue cryopreserved with an elevated thawing rate. Ovarian tissue samples were collected and cryopreserved (frozen and thawed) following RNA sequencing and histological evaluation. Three groups were formed: fresh tissue (Group 1), frozen tissue after quick thawing at 100 \u00b0C (Group 2), and frozen tissue after slow thawing at 37 \u00b0C (Group 3). KEGG analysis showed that in comparison with Group 1, DEGs in Group 2 were mainly enriched in the cortisol synthesis and ovarian steroidogenesis pathways, and DEGs in the cells of Group 3 were mainly enriched in the ovarian steroidogenesis pathway. GO analysis showed that compared to cells of Group 2, DEGs in Group 3 were primarily enriched in the SRP-dependent co-translational protein targeting pathway and co-translational protein targeting to the membrane. The results were formulated with a minimal difference in the histological evaluation of cells after quick and slow thawed tissue. Cryopreservation of ovarian tissue by the described method does not decrease follicle production but downregulates the ovarian steroidogenesis pathway, reducing estrogen and progesterone secretion. The quick thawing of ovarian tissue increases the proliferation and apoptosis pathways of cells.<\/jats:p>","DOI":"10.3390\/ijms252413747","type":"journal-article","created":{"date-parts":[[2024,12,24]],"date-time":"2024-12-24T02:50:56Z","timestamp":1735008656000},"page":"13747","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Positive Effect of Elevated Thawing Rate for Cryopreservation of Human Ovarian Tissue: Transcriptomic Analysis of Fresh and Cryopreserved Cells"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-8398-474X","authenticated-orcid":false,"given":"Qingduo","family":"Kong","sequence":"first","affiliation":[{"name":"Department of Obstetrics and Gynecology, Medical Faculty, Cologne University, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3731-6116","authenticated-orcid":false,"given":"Plamen","family":"Todorov","sequence":"additional","affiliation":[{"name":"Institute of Biology and Immunology of Reproduction of Bulgarian Academy of Sciences (BAS), 1113 Sofia, Bulgaria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cheng","family":"Pei","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Medical Faculty, Cologne University, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Evgenia","family":"Isachenko","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Medical Faculty, Cologne University, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gohar","family":"Rahimi","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Medical Faculty, Cologne University, 50931 Cologne, Germany"},{"name":"Medizinisches Versorgungszentrum AMEDES f\u00fcr IVF- und Pr\u00e4natalmedizin in K\u00f6ln GmbH, 50968 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0781-028X","authenticated-orcid":false,"given":"Nina","family":"Mallmann-Gottschalk","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Medical Faculty, Essen University, 45147 Essen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3674-543X","authenticated-orcid":false,"given":"Volodimir","family":"Isachenko","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Medical Faculty, Cologne University, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,12,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1994","DOI":"10.1200\/JCO.2018.78.1914","article-title":"Fertility preservation in patients with cancer: ASCO clinical practice guideline update","volume":"36","author":"Oktay","year":"2018","journal-title":"J. Clin. Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1016\/j.fertnstert.2019.06.029","article-title":"Advances in fertility-preservation surgery: Navigating new frontiers","volume":"112","author":"Christianson","year":"2019","journal-title":"Fertil. Steril."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"101115","DOI":"10.1016\/j.blre.2023.101115","article-title":"Preservation of fertility in female patients with hematologic diseases","volume":"62","author":"Antunes","year":"2023","journal-title":"Blood Rev."},{"key":"ref_4","first-page":"919","article-title":"Importance of ovarian tissue cryopreservation in fertility preservation and anti-aging treatment","volume":"35","author":"Kasum","year":"2019","journal-title":"Gynecol. Endocrinol. Off. J. Int. Soc. Gynecol. Endocrinol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"489","DOI":"10.1093\/humrep\/deac249","article-title":"Live birth rate after female fertility preservation for cancer or haematopoietic stem cell transplantation: A systematic review and meta-analysis of the three main techniques; embryo, oocyte and ovarian tissue cryopreservation","volume":"38","author":"Fraison","year":"2023","journal-title":"Hum. Reprod."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"777","DOI":"10.1016\/j.molmed.2020.11.003","article-title":"Fertility Preservation: The Challenge of Freezing and Transplanting Ovarian Tissue","volume":"27","author":"Dolmans","year":"2021","journal-title":"Trends Mol. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1016\/j.bpobgyn.2020.06.011","article-title":"Fertility preservation in women for medical and social reasons: Oocytes vs ovarian tissue","volume":"70","author":"Dolmans","year":"2021","journal-title":"Best Pract. Res. Clin. Obstet. Gynaecol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1787.e1","DOI":"10.1016\/j.fertnstert.2010.11.041","article-title":"Pregnancy and live birth after autotransplantation of frozen-thawed ovarian tissue in a patient with metastatic disease undergoing chemotherapy and hematopoietic stem cell transplantation","volume":"95","author":"Donnez","year":"2011","journal-title":"Fertil. Steril."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2413.e15","DOI":"10.1016\/j.fertnstert.2009.12.022","article-title":"Live birth after ovarian tissue autograft in a patient with sickle cell disease treated by allogeneic bone marrow transplantation","volume":"93","author":"Roux","year":"2010","journal-title":"Fertil. Steril."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e4095","DOI":"10.1097\/MD.0000000000004095","article-title":"Comparison of vitrification and conventional slow freezing for cryopreservation of ovarian tissue with respect to the number of intact primordial follicles: A meta-analysis","volume":"95","author":"Zhou","year":"2016","journal-title":"Medicine"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3291","DOI":"10.1007\/s43032-021-00591-6","article-title":"Methods of Ovarian Tissue Cryopreservation: Is Vitrification Superior to Slow Freezing?-Ovarian Tissue Freezing Methods","volume":"28","author":"Kometas","year":"2021","journal-title":"Reprod. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1016\/j.fertnstert.2013.11.016","article-title":"Improving ovarian tissue cryopreservation for oncologic patients: Slow freezing versus vitrification, effect of different procedures and devices","volume":"101","author":"Herraiz","year":"2014","journal-title":"Fertil. Steril."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"841","DOI":"10.1080\/14740338.2019.1636964","article-title":"Safety of fertility treatments in women with systemic autoimmune diseases (SADs)","volume":"18","author":"Vanni","year":"2019","journal-title":"Expert Opin. Drug Saf."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1055\/s-0032-1327812","article-title":"Cryopreservation of Ovarian Tissue: Detailed Description of Methods for Transport, Freezing and Thawing","volume":"72","author":"Isachenko","year":"2012","journal-title":"Geburtshilfe Frauenheilkd"},{"key":"ref_15","first-page":"300","article-title":"Apoptosis in human ovarian tissue after conventional freezing or vitrification and xenotransplantation","volume":"30","author":"Rahimi","year":"2009","journal-title":"CryoLetters"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1016\/j.rbmo.2011.04.005","article-title":"Vitrification as an alternative means of cryopreserving ovarian tissue","volume":"23","author":"Amorim","year":"2011","journal-title":"Reprod. Biomed. Online"},{"key":"ref_17","first-page":"8","article-title":"Retransplantation of cryopreserved ovarian tissue: The first live birth in Germany","volume":"109","author":"Keller","year":"2012","journal-title":"Dtsch. \u00c4rzteblatt Int."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"568","DOI":"10.1038\/340568a0","article-title":"Leucine zippers of fos, jun and GCN4 dictate dimerization specificity and thereby control DNA binding","volume":"340","author":"Kouzarides","year":"1989","journal-title":"Nature"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Gazon, H., Barbeau, B., Mesnard, J.M., and Peloponese, J.M. (2017). Hijacking of the AP-1 Signaling Pathway during Development of ATL. Front. Microbiol., 8.","DOI":"10.3389\/fmicb.2017.02686"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1007\/BF02533989","article-title":"Calmodulin antagonists suppress cholesterol synthesis by inhibiting sterol delta 24 reductase","volume":"22","author":"Filipovic","year":"1987","journal-title":"Lipids"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1042\/bj20011777","article-title":"Inhibition of cholesterol biosynthesis by Delta22-unsaturated phytosterols via competitive inhibition of sterol Delta24-reductase in mammalian cells","volume":"366","author":"Ferruelo","year":"2002","journal-title":"Biochem. J."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1038\/s41586-022-05164-4","article-title":"A phosphoinositide signalling pathway mediates rapid lysosomal repair","volume":"609","author":"Tan","year":"2022","journal-title":"Nature"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1111\/tra.12613","article-title":"Lysosomal membrane permeabilization and cell death","volume":"19","author":"Wang","year":"2018","journal-title":"Traffic"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Yang, C., and Wang, X. (2021). Lysosome biogenesis: Regulation and functions. J. Cell Biol., 220.","DOI":"10.1083\/jcb.202102001"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.cell.2018.09.048","article-title":"Biological Functions of Autophagy Genes: A Disease Perspective","volume":"176","author":"Levine","year":"2019","journal-title":"Cell"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2078","DOI":"10.1080\/15548627.2023.2170962","article-title":"Blockage of autophagosome-lysosome fusion through SNAP29 O-GlcNAcylation promotes apoptosis via ROS production","volume":"19","author":"Pellegrini","year":"2023","journal-title":"Autophagy"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Li, L., Shi, X., Shi, Y., and Wang, Z. (2021). The Signaling Pathways Involved in Ovarian Follicle Development. Front. Physiol., 12.","DOI":"10.3389\/fphys.2021.730196"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1023\/A:1012748718150","article-title":"Multiple signal transduction pathways regulate ovarian steroidogenesis","volume":"3","author":"Wood","year":"2002","journal-title":"Rev. Endocr. Metab. Disord."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"314","DOI":"10.54680\/fr23610110112","article-title":"The technology in cryotechnology","volume":"44","author":"Hagedorn","year":"2023","journal-title":"CryoLetters"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"963689721999617","DOI":"10.1177\/0963689721999617","article-title":"Cryopreservation: An Overview of Principles and Cell-Specific Considerations","volume":"30","author":"Whaley","year":"2021","journal-title":"Cell Transplant."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1007\/978-1-59745-362-2_3","article-title":"Principles of cryopreservation","volume":"368","author":"Pegg","year":"2007","journal-title":"Methods Mol. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1007\/s11626-017-0201-y","article-title":"Best practices for cryopreserving, thawing, recovering, and assessing cells","volume":"53","author":"Baust","year":"2017","journal-title":"Vitr. Cell. Dev. Biol. Anim."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1016\/j.bcmd.2007.06.006","article-title":"Gadd45 in the response of hematopoietic cells to genotoxic stress","volume":"39","author":"Liebermann","year":"2007","journal-title":"Blood Cells Mol. Dis."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1375","DOI":"10.1002\/med.22015","article-title":"GADD45A: With or without you","volume":"44","author":"Palomer","year":"2024","journal-title":"Med. Res. Rev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1002\/jcp.21582","article-title":"Gadd45 modulation of intrinsic and extrinsic stress responses in myeloid cells","volume":"218","author":"Hoffman","year":"2009","journal-title":"J. Cell Physiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/j.ctrv.2003.07.007","article-title":"PI3K\/Akt signalling pathway and cancer","volume":"30","author":"Casado","year":"2004","journal-title":"Cancer Treat. Rev."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2373","DOI":"10.1002\/jcp.27262","article-title":"PI3k\/AKT signaling pathway: Erythropoiesis and beyond","volume":"234","author":"Jafari","year":"2019","journal-title":"J. Cell Physiol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Horta, C.A., Doan, K., and Yang, J. (2023). Mechanotransduction pathways in regulating epithelial-mesenchymal plasticity. Curr. Opin. Cell Biol., 85.","DOI":"10.1016\/j.ceb.2023.102245"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2636","DOI":"10.1111\/febs.15282","article-title":"The ECM path of senescence in aging: Components and modifiers","volume":"287","author":"Levi","year":"2020","journal-title":"FEBS J."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1002\/jcp.10471","article-title":"From cell-ECM interactions to tissue engineering","volume":"199","author":"Rosso","year":"2004","journal-title":"J. Cell Physiol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Jung, S.J., and Kim, H. (2021). Emerging View on the Molecular Functions of Sec62 and Sec63 in Protein Translocation. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222312757"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"736","DOI":"10.2174\/138920311798841717","article-title":"Mechanisms of insertion of tail-anchored proteins into the membrane of the endoplasmic reticulum","volume":"12","author":"Colombo","year":"2011","journal-title":"Curr. Protein Pept. Sci."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1916","DOI":"10.1093\/plphys\/kiab298","article-title":"Looking for a safe haven: Tail-anchored proteins and their membrane insertion pathways","volume":"187","author":"Mehlhorn","year":"2021","journal-title":"Plant Physiol."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Zhou, Y., Wang, W., Todorov, P., Pei, C., Isachenko, E., Rahimi, G., Mallmann, P., Nawroth, F., and Isachenko, V. (2023). RNA Transcripts in Human Ovarian Cells: Two-Time Cryopreservation Does Not Affect Developmental Potential. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24086880"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1186\/s13048-020-00713-9","article-title":"Patient with ovarian insufficiency: Baby born after anticancer therapy and re-transplantation of cryopreserved ovarian tissue","volume":"13","author":"Isachenko","year":"2020","journal-title":"J. Ovarian Res."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Isachenko, V., Mallmann, P., Petrunkina, A.M., Rahimi, G., Nawroth, F., Hancke, K., Felberbaum, R., Genze, F., Damjanoski, I., and Isachenko, E. (2012). Comparison of in vitro- and chorioallantoic membrane (CAM)-culture systems for cryopreserved medulla-contained human ovarian tissue. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0032549"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Isachenko, V., Todorov, P., Isachenko, E., Rahimi, G., Hanstein, B., Salama, M., Mallmann, P., Tchorbanov, A., Hardiman, P., and Getreu, N. (2016). Cryopreservation and xenografting of human ovarian fragments: Medulla decreases the phosphatidylserine translocation rate. Reprod. Biol. Endocrinol. 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