Effects of vitrification and cryostorage duration on single-cell RNA-Seq profiling of vitrified-thawed human metaphase II oocytes
Ying Huo1,2,3,4, Peng Yuan1,3,4, Qingyuan Qin1,3,4, Zhiqiang Yan1,3,4,5, Liying Yan1,3,4,6, Ping Liu1,3,4, Rong Li1,3,4,6, Jie Yan1,3,4(), Jie Qiao1,3,4,5,6
1. Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100191, China 2. Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Peking University, Beijing 100191, China 3. Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproduction, Beijing 100191, China 4. Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing 100191, China 5. Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China 6. National Clinical Research Center of Obstetrics and Gynecology, Beijing 100191, China
Oocyte cryopreservation is widely used for clinical and social reasons. Previous studies have demonstrated that conventional slow-freezing cryopreservation procedures, but not storage time, can alter the gene expression profiles of frozen oocytes. Whether vitrification procedures and the related frozen storage durations have any effects on the transcriptomes of human metaphase II oocytes remain unknown. Four women (30–32 years old) who had undergone IVF treatment were recruited for this study. RNA-Seq profiles of 3 fresh oocytes and 13 surviving vitrified-thawed oocytes (3, 3, 4, and 3 oocytes were cryostored for 1, 2, 3, and 12 months) were analyzed at a single-cell resolution. A total of 1987 genes were differentially expressed in the 13 vitrified-thawed oocytes. However, no differentially expressed genes were found between any two groups among the 1-, 2-, 3-, and 12-month storage groups. Further analysis revealed that the aberrant genes in the vitrified oocytes were closely related to oogenesis and development. Our findings indicated that the effects of vitrification on the transcriptomes of mature human oocytes are induced by the procedure itself, suggesting that long-term cryostorage of human oocytes is safe.
. [J]. Frontiers of Medicine, 2021, 15(1): 144-154.
Ying Huo, Peng Yuan, Qingyuan Qin, Zhiqiang Yan, Liying Yan, Ping Liu, Rong Li, Jie Yan, Jie Qiao. Effects of vitrification and cryostorage duration on single-cell RNA-Seq profiling of vitrified-thawed human metaphase II oocytes. Front. Med., 2021, 15(1): 144-154.
E Stehlik, J Stehlik, KP Katayama, M Kuwayama, V Jambor, R Brohammer, O Kato. Vitrification demonstrates significant improvement versus slow freezing of human blastocysts. Reprod Biomed Online 2005; 11(1): 53–57 https://doi.org/10.1016/S1472-6483(10)61298-9
pmid: 16102287
6
GD Smith, PC Serafini, J Fioravanti, I Yadid, M Coslovsky, P Hassun, JR Alegretti, EL Motta. Prospective randomized comparison of human oocyte cryopreservation with slow-rate freezing or vitrification. Fertil Steril 2010; 94(6): 2088–2095 https://doi.org/10.1016/j.fertnstert.2009.12.065
pmid: 20171613
7
PE Levi Setti, E Porcu, P Patrizio, V Vigiliano, R de Luca, P d’Aloja, R Spoletini, G Scaravelli. Human oocyte cryopreservation with slow freezing versus vitrification. Results from the National Italian Registry data, 2007–2011. Fertil Steril 2014; 102(1): 90–95.e2 https://doi.org/10.1016/j.fertnstert.2014.03.052
pmid: 24794316
8
S Stigliani, S Moretti, P Anserini, I Casciano, PL Venturini, P Scaruffi. Storage time does not modify the gene expression profile of cryopreserved human metaphase II oocytes. Hum Reprod 2015; 30(11): 2519–2526 https://doi.org/10.1093/humrep/dev232
pmid: 26385790
9
L Kuleshova, L Gianaroli, C Magli, A Ferraretti, A Trounson. Birth following vitrification of a small number of human oocytes: case report. Hum Reprod 1999; 14(12): 3077–3079 https://doi.org/10.1093/humrep/14.12.3077
pmid: 10601099
10
L Zhang, X Xue, J Yan, LY Yan, XH Jin, XH Zhu, ZZ He, J Liu, R Li, J Qiao. L-proline: a highly effective cryoprotectant for mouse oocyte vitrification. Sci Rep 2016; 6(1): 26326 https://doi.org/10.1038/srep26326
pmid: 27412080
DA Gook, SM Osborn, WI Johnston. Cryopreservation of mouse and human oocytes using 1,2-propanediol and the configuration of the meiotic spindle. Hum Reprod 1993; 8(7): 1101–1109 https://doi.org/10.1093/oxfordjournals.humrep.a138201
pmid: 8408494
13
MT Zenzes, R Bielecki, RF Casper, SP Leibo. Effects of chilling to 0 °C on the morphology of meiotic spindles in human metaphase II oocytes. Fertil Steril 2001; 75(4): 769–777 https://doi.org/10.1016/S0015-0282(00)01800-8
pmid: 11287033
14
L Rienzi, F Martinez, F Ubaldi, MG Minasi, M Iacobelli, J Tesarik, E Greco. Polscope analysis of meiotic spindle changes in living metaphase II human oocytes during the freezing and thawing procedures. Hum Reprod 2004; 19(3): 655–659 https://doi.org/10.1093/humrep/deh101
pmid: 14998966
15
Y Ghetler, E Skutelsky, I Ben Nun, L Ben Dor, D Amihai, R Shalgi. Human oocyte cryopreservation and the fate of cortical granules. Fertil Steril 2006; 86(1): 210–216 https://doi.org/10.1016/j.fertnstert.2005.12.061
pmid: 16756978
16
V Bianchi, G Macchiarelli, A Borini, M Lappi, S Cecconi, S Miglietta, G Familiari, SA Nottola. Fine morphological assessment of quality of human mature oocytes after slow freezing or vitrification with a closed device: a comparative analysis. Reprod Biol Endocrinol 2014; 12(1): 110 https://doi.org/10.1186/1477-7827-12-110
pmid: 25421073
17
S Succu, D Bebbere, L Bogliolo, F Ariu, S Fois, GG Leoni, F Berlinguer, S Naitana, S Ledda. Vitrification of in vitro matured ovine oocytes affects in vitro pre-implantation development and mRNA abundance. Mol Reprod Dev 2008; 75(3): 538–546 https://doi.org/10.1002/mrd.20784
pmid: 17886274
18
VM Anchamparuthy, RE Pearson, FC Gwazdauskas. Expression pattern of apoptotic genes in vitrified-thawed bovine oocytes. Reprod Domest Anim 2010; 45(5): e83–e90
pmid: 19821945
19
B Turathum, K Saikhun, P Sangsuwan, Y Kitiyanant. Effects of vitrification on nuclear maturation, ultrastructural changes and gene expression of canine oocytes. Reprod Biol Endocrinol 2010; 8(1): 70 https://doi.org/10.1186/1477-7827-8-70
pmid: 20565987
20
A Habibi, N Farrokhi, F Moreira da Silva, BF Bettencourt, J Bruges-Armas, F Amidi, A Hosseini. The effects of vitrification on gene expression in mature mouse oocytes by nested quantitative PCR. J Assist Reprod Genet 2010; 27(11): 599–604 https://doi.org/10.1007/s10815-010-9453-0
pmid: 20714800
21
C Di Pietro, M Vento, MR Guglielmino, P Borzì, M Santonocito, M Ragusa, D Barbagallo, LR Duro, A Majorana, A De Palma, MR Garofalo, E Minutolo, P Scollo, M Purrello. Molecular profiling of human oocytes after vitrification strongly suggests that they are biologically comparable with freshly isolated gametes. Fertil Steril 2010; 94(7): 2804–2807 https://doi.org/10.1016/j.fertnstert.2010.04.060
pmid: 20542504
22
C Monzo, D Haouzi, K Roman, S Assou, H Dechaud, S Hamamah. Slow freezing and vitrification differentially modify the gene expression profile of human metaphase II oocytes. Hum Reprod 2012; 27(7): 2160–2168 https://doi.org/10.1093/humrep/des153
pmid: 22587994
23
S Chamayou, G Bonaventura, C Alecci, D Tibullo, F Di Raimondo, A Guglielmino, ML Barcellona. Consequences of metaphase II oocyte cryopreservation on mRNA content. Cryobiology 2011; 62(2): 130–134 https://doi.org/10.1016/j.cryobiol.2011.01.014
pmid: 21272569
24
MG Larman, MG Katz-Jaffe, CB Sheehan, DK Gardner. 1,2-propanediol and the type of cryopreservation procedure adversely affect mouse oocyte physiology. Hum Reprod 2007; 22(1): 250–259 https://doi.org/10.1093/humrep/del319
pmid: 16905767
25
MG Katz-Jaffe, MG Larman, CB Sheehan, DK Gardner. Exposure of mouse oocytes to 1,2-propanediol during slow freezing alters the proteome. Fertil Steril 2008; 89(5 Suppl): 1441–1447 https://doi.org/10.1016/j.fertnstert.2007.03.098
pmid: 17980362
A Borini, G Coticchio. The efficacy and safety of human oocyte cryopreservation by slow cooling. Semin Reprod Med 2009; 27(6): 443–449 https://doi.org/10.1055/s-0029-1241053
pmid: 19806512
28
F Ubaldi, R Anniballo, S Romano, E Baroni, L Albricci, S Colamaria, A Capalbo, F Sapienza, G Vajta, L Rienzi. Cumulative ongoing pregnancy rate achieved with oocyte vitrification and cleavage stage transfer without embryo selection in a standard infertility program. Hum Reprod 2010; 25(5): 1199–1205 https://doi.org/10.1093/humrep/deq046
pmid: 20185513
29
A Cobo, C Diaz. Clinical application of oocyte vitrification: a systematic review and meta-analysis of randomized controlled trials. Fertil Steril 2011; 96(2): 277–285 https://doi.org/10.1016/j.fertnstert.2011.06.030
pmid: 21718983
30
L Rienzi, A Cobo, A Paffoni, C Scarduelli, A Capalbo, G Vajta, J Remohí, G Ragni, FM Ubaldi. Consistent and predictable delivery rates after oocyte vitrification: an observational longitudinal cohort multicentric study. Hum Reprod 2012; 27(6): 1606–1612 https://doi.org/10.1093/humrep/des088
pmid: 22442248
31
L Yan, M Yang, H Guo, L Yang, J Wu, R Li, P Liu, Y Lian, X Zheng, J Yan, J Huang, M Li, X Wu, L Wen, K Lao, R Li, J Qiao, F Tang. Single-cell RNA-Seq profiling of human preimplantation embryos and embryonic stem cells. Nat Struct Mol Biol 2013; 20(9): 1131–1139 https://doi.org/10.1038/nsmb.2660
pmid: 23934149
32
F Tang, C Barbacioru, E Nordman, B Li, N Xu, VI Bashkirov, K Lao, MA Surani. RNA-Seq analysis to capture the transcriptome landscape of a single cell. Nat Protoc 2010; 5(3): 516–535 https://doi.org/10.1038/nprot.2009.236
pmid: 20203668
33
F Zhou, X Li, W Wang, P Zhu, J Zhou, W He, M Ding, F Xiong, X Zheng, Z Li, Y Ni, X Mu, L Wen, T Cheng, Y Lan, W Yuan, F Tang, B Liu. Tracing haematopoietic stem cell formation at single-cell resolution. Nature 2016; 533(7604): 487–492 https://doi.org/10.1038/nature17997
pmid: 27225119
34
YX Cao, Q Xing, L Li, L Cong, ZG Zhang, ZL Wei, P Zhou. Comparison of survival and embryonic development in human oocytes cryopreserved by slow-freezing and vitrification. Fertil Steril 2009; 92(4): 1306–1311 https://doi.org/10.1016/j.fertnstert.2008.08.069
pmid: 18930218
35
R Fadini, F Brambillasca, MM Renzini, M Merola, R Comi, E De Ponti, MB Dal Canto. Human oocyte cryopreservation: comparison between slow and ultrarapid methods. Reprod Biomed Online 2009; 19(2): 171–180 https://doi.org/10.1016/S1472-6483(10)60069-7
pmid: 19712551
36
DH Edgar, DA Gook. A critical appraisal of cryopreservation (slow cooling versus vitrification) of human oocytes and embryos. Hum Reprod Update 2012; 18(5): 536–554 https://doi.org/10.1093/humupd/dms016
pmid: 22537859
37
L Parmegiani, C Garello, F Granella, D Guidetti, S Bernardi, GE Cognigni, A Revelli, M Filicori. Long-term cryostorage does not adversely affect the outcome of oocyte thawing cycles. Reprod Biomed Online 2009; 19(3): 374–379 https://doi.org/10.1016/S1472-6483(10)60171-X
pmid: 19778482
38
KN Goldman, Y Kramer, B Hodes-Wertz, N Noyes, C McCaffrey, JA Grifo. Long-term cryopreservation of human oocytes does not increase embryonic aneuploidy. Fertil Steril 2015; 103(3): 662–668 https://doi.org/10.1016/j.fertnstert.2014.11.025
pmid: 25542819
39
SB Qian, H McDonough, F Boellmann, DM Cyr, C Patterson. CHIP-mediated stress recovery by sequential ubiquitination of substrates and Hsp70. Nature 2006; 440(7083): 551–555 https://doi.org/10.1038/nature04600
pmid: 16554822
40
YJ Chen, KC Lai, HH Kuo, LP Chow, LH Yih, TC Lee. HSP70 colocalizes with PLK1 at the centrosome and disturbs spindle dynamics in cells arrested in mitosis by arsenic trioxide. Arch Toxicol 2014; 88(9): 1711–1723 https://doi.org/10.1007/s00204-014-1222-x
pmid: 24623308
A Kishor, EJF White, AE Matsangos, Z Yan, B Tandukar, GM Wilson. Hsp70’s RNA-binding and mRNA-stabilizing activities are independent of its protein chaperone functions. J Biol Chem 2017; 292(34): 14122–14133 https://doi.org/10.1074/jbc.M117.785394
pmid: 28679534
43
M Mišunová, T Svitálková, L Pleštilová, O Kryštufková, D Tegzová, R Svobodová, M Hušáková, M Tomčík, R Bečvář, J Závada, H Mann, L Kolesár, A Slavčev, J Vencovský, P Novota. Molecular markers of systemic autoimmune disorders: the expression of MHC-located HSP70 genes is significantly associated with autoimmunity development. Clin Exp Rheumatol 2017; 35(1): 33–42
pmid: 28032847
44
S Takahashi, G Andreoletti, R Chen, Y Munehira, A Batra, NA Afzal, RM Beattie, JA Bernstein, S Ennis, M Snyder. De novo and rare mutations in the HSPA1L heat shock gene associated with inflammatory bowel disease. Genome Med 2017; 9(1): 8 https://doi.org/10.1186/s13073-016-0394-9
pmid: 28126021
45
F Le Masson, E Christians. HSFs and regulation of Hsp70.1 (Hspa1b) in oocytes and preimplantation embryos: new insights brought by transgenic and knockout mouse models. Cell Stress Chaperones 2011; 16(3): 275–285 https://doi.org/10.1007/s12192-010-0239-1
pmid: 21053113
46
JM Lelièvre, N Peynot, S Ruffini, L Laffont, D Le Bourhis, PM Girard, V Duranthon. Regulation of heat-inducible HSPA1A gene expression during maternal-to-embryo transition and in response to heat in in vitro-produced bovine embryos. Reprod Fertil Dev 2017; 29(9): 1868–1881 https://doi.org/10.1071/RD15504
pmid: 27851888
47
C Schumacher, H Wang, C Honer, W Ding, J Koehn, Q Lawrence, CM Coulis, LL Wang, D Ballinger, BR Bowen, S Wagner. The SCAN domain mediates selective oligomerization. J Biol Chem 2000; 275(22): 17173–17179 https://doi.org/10.1074/jbc.M000119200
pmid: 10747874
FR Carneiro, TC Silva, AC Alves, T Haline-Vaz, FC Gozzo, NI Zanchin. Spectroscopic characterization of the tumor antigen NY-REN-21 and identification of heterodimer formation with SCAND1. Biochem Biophys Res Commun 2006; 343(1): 260–268 https://doi.org/10.1016/j.bbrc.2006.02.140
pmid: 16540086
50
T Oka, M Krieger. Multi-component protein complexes and Golgi membrane trafficking. J Biochem 2005; 137(2): 109–114 https://doi.org/10.1093/jb/mvi024
pmid: 15749823
51
I Martianov, A Ramadass, A Serra Barros, N Chow, A Akoulitchev. Repression of the human dihydrofolate reductase gene by a non-coding interfering transcript. Nature 2007; 445(7128): 666–670 https://doi.org/10.1038/nature05519
pmid: 17237763
52
JL Rinn, M Kertesz, JK Wang, SL Squazzo, X Xu, SA Brugmann, LH Goodnough, JA Helms, PJ Farnham, E Segal, HY Chang. Functional demarcation of active and silent chromatin domains in human HOX loci by noncoding RNAs. Cell 2007; 129(7): 1311–1323 https://doi.org/10.1016/j.cell.2007.05.022
pmid: 17604720
53
AM Khalil, M Guttman, M Huarte, M Garber, A Raj, D Rivea Morales, K Thomas, A Presser, BE Bernstein, A van Oudenaarden, A Regev, ES Lander, JL Rinn. Many human large intergenic noncoding RNAs associate with chromatin-modifying complexes and affect gene expression. Proc Natl Acad Sci USA 2009; 106(28): 11667–11672 https://doi.org/10.1073/pnas.0904715106
pmid: 19571010
54
S Loewer, MN Cabili, M Guttman, YH Loh, K Thomas, IH Park, M Garber, M Curran, T Onder, S Agarwal, PD Manos, S Datta, ES Lander, TM Schlaeger, GQ Daley, JL Rinn. Large intergenic non-coding RNA-RoR modulates reprogramming of human induced pluripotent stem cells. Nat Genet 2010; 42(12): 1113–1117 https://doi.org/10.1038/ng.710
pmid: 21057500
55
J Li, Z Gao, X Wang, H Liu, Y Zhang, Z Liu. Identification and functional analysis of long intergenic noncoding RNA genes in porcine pre-implantation embryonic development. Sci Rep 2016; 6(1): 38333 https://doi.org/10.1038/srep38333
pmid: 27922056
56
M Guttman, J Donaghey, BW Carey, M Garber, JK Grenier, G Munson, G Young, AB Lucas, R Ach, L Bruhn, X Yang, I Amit, A Meissner, A Regev, JL Rinn, DE Root, ES Lander. lincRNAs act in the circuitry controlling pluripotency and differentiation. Nature 2011; 477(7364): 295–300 https://doi.org/10.1038/nature10398
pmid: 21874018
57
M Guttman, I Amit, M Garber, C French, MF Lin, D Feldser, M Huarte, O Zuk, BW Carey, JP Cassady, MN Cabili, R Jaenisch, TS Mikkelsen, T Jacks, N Hacohen, BE Bernstein, M Kellis, A Regev, JL Rinn, ES Lander. Chromatin signature reveals over a thousand highly conserved large non-coding RNAs in mammals. Nature 2009; 458(7235): 223–227 https://doi.org/10.1038/nature07672
pmid: 19182780