Impact of embryo density on the proportion of good quality cleavage embryos

SHI Cheng; CHEN Shu-Wen; WANG Ping; LIANG Rong; DUAN Sheng-Nan; CHEN Xi

doi:10.16098/j.issn.0529-1356.2021.01.022

PDF(2574 KB)

Acta Anatomica Sinica ›› 2021, Vol. 52 ›› Issue (1) : 135-140. DOI: 10.16098/j.issn.0529-1356.2021.01.022

Histology,Embryology and Developmental Biology

Impact of embryo density on the proportion of good quality cleavage embryos

SHI Cheng CHEN Shu-wen WANG Ping LIANG Rong DUAN Sheng-nan CHEN Xi*

Author information +

History +

Abstract

Objective To investigate the real correlation between embryo density and developmental outcome of cleavage embryos on day 3 by adjusting covariates. Methods Data of 1196 embryos from 206 couples undergoing in vitro fertilization and embryo transfer(IVF-ET) treatment were retrospectively analyzed. Embryos were hypoxia-cultivated in a fixed 30 μl microdrop in culture dishes. Embryo quality on day 3 was evaluated and proportion of good quality cleavage embryos on day 3 was used as the endpoint. Maternal age, paternal age, antral follicles, level of anti-mullerian hormone (AMH), type of infertility, controlled ovarian stimulation(COS) protocol, length of stimulation, number of retrieved oocytes, and type of insemination were chosen as covariates. Results A total of 1196 embryos were included and analyzed. Three embryo densities were routinely used: 30 μl/embryo ［1 embryo/(30 μl·drop)］, 15 μl/embryo ［2 embryos /(30 μl·drop)］ and 10 μl/embryo ［3 embryos/(30 μl·drop)］. The number of embryos assigned into these three groups were 434, 646 and 116 embryos separately. The average proportion of good quality embryos on day 3 in 10 μl/embryo group were lower than that in both 15 and 30 μl/embryo group (29.31% vs 40.25%，P<0.05；29.31% vs 42.17%，P<0.05), and it was comparable between groups of 15 and 30 μl/embryo (40.25% vs 42.17%，P>0.05). In the regression analysis, without adjusting any covariables, the good quality embryos rate on day 3 of the 10 μl/embryo group was 43% lower than that of the 30 μl/embryo group (0.57, 95% CI 0.32, 1.03), and there was no statistical difference (P>0.05). In the minimum-adjusted model 2 (adjusted the level of AMH and type of insemination), proportion of good quality embryos on day 3 in group of 10 μl/embryo significantly decreased by 51% (0.49, 95%CI 0.27，0.90, P<0.05) compared with that in group of 30 μl/embryo.

ConclusionIn a 30 μl microdrop, compared with individual cul turing, group culturing with the density of 10 μl/embryo did not benefit the development of the cleavage embryos and 30 μl/embryo was the optimal embryo density.

Key words

Embryo density / Embryo quality / Individual culture / Group culture / Embryo culture / Female

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

SHI Cheng CHEN Shu-wen WANG Ping LIANG Rong DUAN Sheng-nan CHEN Xi. Impact of embryo density on the proportion of good quality cleavage embryos[J]. Acta Anatomica Sinica. 2021, 52(1): 135-140 https://doi.org/10.16098/j.issn.0529-1356.2021.01.022

References

［1］ Lehner A, Kaszas Z, Murber A, et al. Embryo density may affect embryo quality during in vitro culture in a microwell group culture dish［J］. Arch Gynecol Obstet，2017, 296(2): 345-353.

［2］ Ebner T, Shebl O, Moser M, et al. Group culture of human zygotes is superior to individual culture in terms of blastulation, implantation and life birth［J］. Reprod Biomed Online，2010, 21(6): 762-768.

［3］ Minasi MG, Fabozzi G, Casciani V, et al. Improved blastocyst formation with reduced culture volume: comparison of three different culture conditions on 1128 sibling human zygotes［J］. J Assist Reprod Genet， 2015, 32(2): 215-220.

［4］ De Munck N, Santos-Ribeiro S, Mateizel Ⅰ, et al. Reduced blastocyst formation in reduced culture volume［J］. J Assist Reprod Genet，2015, 32(9): 1365-1370.

［5］ Rebollar-Lazaro I, Matson P. The culture of human cleavage stage embryos alone or in groups: effect upon blastocyst utilization rates and implantation［J］. Reprod Biol，2010, 10(3): 227-234.

［6］ Schini SA, Bavister BD. Development of golden hamster embryos through the two-cell block in chemically defined medium［J］. J Exp Zool，1988, 245(1): 111-115.

［7］ Salahuddin S, Ookutsu S, Goto K, et al. Effects of embryo density and co-culture of unfertilized oocytes on embryonic development of in-vitro fertilized mouse embryos［J］. Hum Reprod，1995, 10(9): 2382-2385.

［8］ Vutyavanich T, Saeng-Anan U, Sirisukkasem S, et al. Effect of embryo density and microdrop volume on the blastocyst development of mouse two-cell embryos［J］. Fertil Steril， 2011, 95(4): 1435-1439.

［9］ O’Doherty EM, Wade MG, Hill JL, et al. Effects of culturing bovine oocytes either singly or in groups on development to blastocysts［J］. Theriogenology，1997, 48(1): 161-169.

［10］ Gardner DK, Lane M, Spitzer A, et al. Enhanced rates of cleavage and development for sheep zygotes cultured to the blastocyst stage in vitro in the absence of serum and somatic cells: amino acids, vitamins, and culturing embryos in groups stimulate development［J］. Biol Reprod，1994, 50(2): 390-400.

［11］ Almagor M, Bejar C, Kafka I, et al. Pregnancy rates after communal growth of preimplantation human embryos in vitro［J］. Fertil Steril, 1996, 66(3): 394-397.

［12］ Moessner J, Dodson WC. The quality of human embryo growth is improved when embryos are cultured in groups rather than separately［J］. Fertiity Steril,1995, 64(5): 1034-1035.

［13］ Christianson MS, Zhao Y, Shoham G, et al. Embryo catheter loading and embryo culture techniques: results of a worldwide web-based survey［J］. J Assist Reprod Genet, 2014, 31(8): 1029-1036.

［14］ Rubio Ⅰ, Galán A, Larreategui Z, et al. Clinical validation of embryo culture and selection by morphokinetic analysis: a randomized, controlled trial of the EmbryoScope［J］. Fertil Steril, 2014, 102(5): 1287-1294.

［15］ Marhuenda-Egea FC, Martínez-Sabater E, Gonsálvez-Alvarez R, et al. A crucial step in assisted reproduction technology: human embryo selection using metabolomic evaluation［J］. Fertil Steril, 2010, 94(2): 772-774.

［16］ Seli E, Vergouw CG, Morita H, et al. Noninvasive metabolomic profiling as an adjunct to morphology for noninvasive embryo assessment in women undergoing single embryo transfer［J］. Fertil Steril, 2010, 94(2): 535-542.

［17］ Leaver M, Wells D. Non-invasive preimplantation genetic testing (niPGT): the next revolution in reproductive genetics［J］? Hum Reprod Update, 2020, 26(1): 16-42.

［18］ Ziebe S, Loft A, Petersen JH, et al. Embryo quality and developmental potential is compromised by age［J］. Acta Obstet Gynecol Scand, 2001, 80(2): 169-174.

［19］ Ji J, Liu Y, Tong XH, et al. The optimum number of oocytes in IVF treatment: an analysis of 2455 cycles in China［J］. Hum Reprod, 2013, 28(10): 2728-2734.

［20］ Kim JY, Kim JH, Jee BC, et al. Can intracytoplasmic sperm injection prevent total fertilization failure and enhance embryo quality in patients with non-male factor infertility ［J］? Eur J Obstet Gynecol Reprod Biol, 2014, 178: 188-191.

［21］ Kuai YR, Wan Sh, Zhang K, et al. Comparing clinical outcome of different methods for cryopreservation of human cleavage stage embryos［J］. Acta Anatomica Sinica, 2017, 48(4): 482-487.（in Chinese）

郐艳荣，王晟，张凯,等. 冷冻方法对人卵裂期冷冻胚胎移植周期妊娠结局的影响［J］. 解剖学报, 2017, 48(4): 482-487.

［22］ Hoelker M, Rings F, Lund Q, et al. Effect of the microenvironment and embryo density on developmental characteristics and gene expression profile of bovine preimplantative embryos cultured in vitro［J］. Reproduction, 2009, 137(3): 415-425.

［23］ Sananmuang T, Tharasanit T, Nguyen C, et al. Culture medium and embryo density influence on developmental competence and gene expression of cat embryos［J］. Theriogenology, 2011, 75(9): 1708-1719.

［24］ Kelley RL, Gardner DK. In vitro culture of individual mouse preimplantation embryos: the role of embryo density, microwells, oxygen, timing and conditioned media［J］. Reprod Biomed Online, 2017, 34(5): 441-454.

Funding

Chinese Medical Association Special Funding Project for Clinical Medical Scientific Research;Application of Clinical Features of Capital Special Subject