An integrated investigation of oocyte developmental competence: expression of key genes in human cumulus cells, morphokinetics of early divisions, blastulation, and euploidy

  • C. Scarica
  • D. Cimadomo
  • L. Dovere
  • A. Giancani
  • M. Stoppa
  • A. Capalbo
  • F. M. Ubaldi
  • L. RienziEmail author
  • R. Canipari
Assisted Reproduction Technologies



To investigate the association of cumulus cell (CC)-related expression of a selected cluster of key genes (PTGS2, CAMK1D, HAS2, STC1, and EFNB2) with embryo development to blastocyst.


Exploratory study at a private clinic. Eighteen advanced maternal age patients were enrolled (37.3 ± 4.0 years). Seventy-five cumuli were collected, whose oocytes resulted in either developmental arrest (N = 33) or blastocyst formation (N = 42). The noninvasive CC gene expression was combined with time-lapse morphokinetic parameters and, for blastocysts, with qPCR-based aneuploidy testing on trophectoderm biopsies.


The detection rate was 100% for all transcripts, but STC1 (96%) and CAMK1D (89%). Among amplified assays, CC mean expression levels of CAMK1D, PTGS2, and HAS2 were higher from oocytes that developed to blastocyst. No difference in CC key gene expression was reported between euploid (N = 21) and aneuploid (N = 21) blastocysts. Some timings of early embryo development were faster in embryos developing to blastocyst (time of pronuclei appearance and fading, division to two- and four-cells, first and second cell cycles). However, the generalized linear models outlined increasing CAMK1D expression levels as the strongest parameter associated with oocytes’ developmental potential from both a general (AUC = 0.78 among amplified samples) and an intrapatient perspectives (AUC = 0.9 among patients obtaining ≥ 2 zygotes from the cohort with different developmental outcomes).


CAMK1D level of expression in CCs associated with blastocyst development. If confirmed from larger studies in wider populations of patients, the investigation of CC key gene expression might suit IVF clinics not adopting blastocyst culture. Future investigations should clarify the role of CAMK1D in ovarian physiology and could provide novel insights on how oocytes gain competence during folliculogenesis.


Oocyte competence Cumulus cells Blastocysts Noninvasive embryo selection CAMK1D 


Authors’ contribution

CS, DC, AC, LR, and RC designed the study. CS and DC collected and analyzed the samples. CS, DC, and RC drafted the manuscript. All authors contributed to the interpretation and discussion of the data.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10815_2019_1410_Fig4_ESM.png (62 kb)
Supplementary Figure 1

Mean Ct values forB2MandUBCin the cumulus cells from oocytes that underwent developmental arrest versus blastocyst formation. Mann-Whitney U tests were conducted that did not show any statistically significant difference. (PNG 62 kb)

10815_2019_1410_MOESM1_ESM.tif (5.4 mb)
High resolution image (TIF 5501 kb)
10815_2019_1410_Fig5_ESM.png (85 kb)
Supplementary Figure 2

Geometric mean ofB2MandUBCCt values in the samples whereSTC1andCAMK1Dwere detected versus the samples where these key-genes were not detected. Mann-Whitney U tests were conducted. Effect size d and post-hoc power analyses were also reported. (PNG 84 kb)

10815_2019_1410_MOESM2_ESM.tif (5 mb)
High resolution image (TIF 5121 kb)
10815_2019_1410_Fig6_ESM.png (68 kb)
Supplementary Figure 3

Relative expression levels ofSTC1andEFNB2from the cumulus cells of developmentally incompetent versus competent oocytes. Total RNA was extracted from cumulus cells obtained from oocytes that resulted in either arrested embryos or blastocysts after IVF. Quantitative-PCR was conducted using Taqman primers. Each reaction was performed in triplicate. Each sample was normalized to its B2M and UBC mRNA content. The mean 2^-dCT values with SD and range are reported next to each boxplot. Mann-Whitney U tests were conducted to assess statistically-significant differences. No statistically-significant difference was defined. Circles and stars represent outliers values (PNG 67 kb)

10815_2019_1410_MOESM3_ESM.tif (4.8 mb)
High resolution image (TIF 4942 kb)
10815_2019_1410_Fig7_ESM.png (76 kb)
Supplementary Figure 4

Relative expression levels ofCAMK1D,PTGS2andHAS2from the cumulus cells whose oocytes developed as aneuploid versus euploid blastocysts (sub-analysis). Total RNA was extracted from cumulus cells obtained from oocytes that resulted in either aneuploid or euploid blastocysts after IVF. Quantitative-PCR was conducted using Taqman primers. Each reaction was performed in triplicate. Each sample was normalized to its B2M and UBC mRNA content. The mean 2^-dCT values with SD and range are reported next to each boxplot. Mann-Whitney U tests were conducted to assess statistically-significant differences. No statistically-significant difference was reported. Effect size d and post-hoc power analyses were also calculated. Circles and stars represent outliers values (PNG 76 kb)

10815_2019_1410_MOESM4_ESM.tif (3.2 mb)
High resolution image (TIF 3275 kb)
10815_2019_1410_Fig8_ESM.png (126 kb)
Supplementary Figure 5

Relative expression levels ofCAMK1D,PTGS2andHAS2from the cumulus cells of oocytes that developed as high quality (class 1&2) and low quality (class 3&4) blastocysts or underwent developmental arrest (sub-analysis). Total RNA was extracted from cumulus cells obtained from oocytes that resulted in either arrested embryos or blastocysts after IVF. Quantitative-PCR was conducted using Taqman primers. Each reaction was performed in triplicate. Each sample was normalized to its B2M and UBC mRNA content. The mean 2^-dCT values with SD and range are reported next to each boxplot. Mann-Whitney U tests were conducted to assess statistically-significant differences. Effect size d and post-hoc power analyses were also reported. Circles and stars represent outliers values (PNG 125 kb)

10815_2019_1410_MOESM5_ESM.tif (4.1 mb)
High resolution image (TIF 4236 kb)
10815_2019_1410_Fig9_ESM.png (90 kb)
Supplementary Figure 6

Patient-specific mean CCs’ expression of selected key-genes with 95%CI entailing all the samples that were amplified within each cohort of zygotes. Kruskal-Wallis tests were conducted to assess statistically significant differences (PNG 89 kb)

10815_2019_1410_MOESM6_ESM.tif (6 mb)
High resolution image (TIF 6171 kb)
10815_2019_1410_Fig10_ESM.png (115 kb)
Supplementary Figure 7

Receiver Operating Characteristic (ROC) curve analysis for the predictive power ofCAMK1D2^-dCT upon oocyte developmental competence in vitro from both a general (a) and a patient-specific (b) analyses conducted from this dataset. The former analysis entailed 67 of the 75 cumuli included in this study (89%), namely the samples where this transcript was detected; the latter analysis instead included the patients (N = 11) who obtained at least two oocytes from the cohort with opposite outcomes (arrested embryo and blastocyst development) and whose cumulus cells resulted in CAMK1D detection. AUC, area under the curve. (PNG 115 kb)

10815_2019_1410_MOESM7_ESM.tif (4.7 mb)
High resolution image (TIF 4831 kb)
10815_2019_1410_MOESM8_ESM.docx (18 kb)
Supplementary Table 1 (DOCX 17 kb)
10815_2019_1410_MOESM9_ESM.docx (31 kb)
Supplementary Table 2 (DOCX 31 kb)


  1. 1.
    Ubaldi FM, Capalbo A, Colamaria S, Ferrero S, Maggiulli R, Vajta G, et al. Reduction of multiple pregnancies in the advanced maternal age population after implementation of an elective single embryo transfer policy coupled with enhanced embryo selection: pre- and post-intervention study. Hum Reprod. 2015;30(9):2097–106. Scholar
  2. 2.
    Guerif F, Le Gouge A, Giraudeau B, Poindron J, Bidault R, Gasnier O, et al. Limited value of morphological assessment at days 1 and 2 to predict blastocyst development potential: a prospective study based on 4042 embryos. Hum Reprod. 2007;22(7):1973–81. Scholar
  3. 3.
    Racowsky C, Ohno-Machado L, Kim J, Biggers JD. Is there an advantage in scoring early embryos on more than one day? Hum Reprod. 2009;24(9):2104–13. Scholar
  4. 4.
    Glujovsky D, Farquhar C. Cleavage-stage or blastocyst transfer: what are the benefits and harms? Fertil Steril. 2016;106(2):244–50. Scholar
  5. 5.
    Gardner DK. Blastocyst culture: toward single embryo transfers. Hum Fertil (Camb). 2000;3(4):229–37.Google Scholar
  6. 6.
    Gardner DK. The impact of physiological oxygen during culture, and vitrification for cryopreservation, on the outcome of extended culture in human IVF. Reprod BioMed Online. 2016;32(2):137–41. Scholar
  7. 7.
    Conaghan J, Chen AA, Willman SP, Ivani K, Chenette PE, Boostanfar R, et al. Improving embryo selection using a computer-automated time-lapse image analysis test plus day 3 morphology: results from a prospective multicenter trial. Fertil Steril. 2013;100(2):412–9 e5. Scholar
  8. 8.
    Kirkegaard K, Kesmodel US, Hindkjaer JJ, Ingerslev HJ. Time-lapse parameters as predictors of blastocyst development and pregnancy outcome in embryos from good prognosis patients: a prospective cohort study. Hum Reprod. 2013;28(10):2643–51. Scholar
  9. 9.
    Meseguer M, Herrero J, Tejera A, Hilligsoe KM, Ramsing NB, Remohi J. The use of morphokinetics as a predictor of embryo implantation. Hum Reprod. 2011;26(10):2658–71. Scholar
  10. 10.
    Armstrong S, Bhide P, Jordan V, Pacey A, Farquhar C. Time-lapse systems for embryo incubation and assessment in assisted reproduction. Cochrane Database Syst Rev. 2018;5:CD011320. Scholar
  11. 11.
    Heffner LJ. Advanced maternal age—how old is too old? N Engl J Med. 2004;351(19):1927–9. Scholar
  12. 12.
    Hassold T, Hunt P. To err (meiotically) is human: the genesis of human aneuploidy. Nat Rev Genet. 2001;2(4):280–91. Scholar
  13. 13.
    Franasiak JM, Forman EJ, Hong KH, Werner MD, Upham KM, Treff NR, et al. The nature of aneuploidy with increasing age of the female partner: a review of 15,169 consecutive trophectoderm biopsies evaluated with comprehensive chromosomal screening. Fertil Steril. 2014;101(3):656–63 e1. Scholar
  14. 14.
    Capalbo A, Hoffmann ER, Cimadomo D, Maria Ubaldi F, Rienzi L. Human female meiosis revised: new insights into the mechanisms of chromosome segregation and aneuploidies from advanced genomics and time-lapse imaging. Hum Reprod Update. 2017;23:1–17. Scholar
  15. 15.
    Dahdouh EM, Balayla J, Garcia-Velasco JA. Comprehensive chromosome screening improves embryo selection: a meta-analysis. Fertil Steril. 2015;104(6):1503–12. Scholar
  16. 16.
    Chen M, Wei S, Hu J, Quan S. Can comprehensive chromosome screening technology improve IVF/ICSI outcomes? A meta-analysis. PLoS One. 2015;10(10):e0140779. Scholar
  17. 17.
    Scott RT Jr, Upham KM, Forman EJ, Zhao T, Treff NR. Cleavage-stage biopsy significantly impairs human embryonic implantation potential while blastocyst biopsy does not: a randomized and paired clinical trial. Fertil Steril. 2013;100(3):624–30. Scholar
  18. 18.
    Cimadomo D, Capalbo A, Ubaldi FM, Scarica C, Palagiano A, Canipari R, et al. The impact of biopsy on human embryo developmental potential during preimplantation genetic diagnosis. Biomed Res Int. 2016;2016:7193075. Scholar
  19. 19.
    Pennetta F, Lagalla C, Borini A. Embryo morphokinetic characteristics and euploidy. Curr Opin Obstet Gynecol. 2018;30(3):185–96. Scholar
  20. 20.
    Campbell A, Fishel S, Bowman N, Duffy S, Sedler M, Hickman CF. Modelling a risk classification of aneuploidy in human embryos using non-invasive morphokinetics. Reprod BioMed Online. 2013;26(5):477–85. Scholar
  21. 21.
    Basile N, Nogales Mdel C, Bronet F, Florensa M, Riqueiros M, Rodrigo L, et al. Increasing the probability of selecting chromosomally normal embryos by time-lapse morphokinetics analysis. Fertil Steril. 2014;101(3):699–704. Scholar
  22. 22.
    Rienzi L, Capalbo A, Stoppa M, Romano S, Maggiulli R, Albricci L, et al. No evidence of association between blastocyst aneuploidy and morphokinetic assessment in a selected population of poor-prognosis patients: a longitudinal cohort study. Reprod BioMed Online. 2015;30(1):57–66. Scholar
  23. 23.
    Eppig JJ. Oocyte control of ovarian follicular development and function in mammals. Reproduction. 2001;122(6):829–38.Google Scholar
  24. 24.
    Paulini F, Melo EO. The role of oocyte-secreted factors GDF9 and BMP15 in follicular development and oogenesis. Reprod Domest Anim. 2011;46(2):354–61. Scholar
  25. 25.
    Trombly DJ, Woodruff TK, Mayo KE. Roles for transforming growth factor beta superfamily proteins in early folliculogenesis. Semin Reprod Med. 2009;27(1):14–23. Scholar
  26. 26.
    Kedem A, Fisch B, Garor R, Ben-Zaken A, Gizunterman T, Felz C, et al. Growth differentiating factor 9 (GDF9) and bone morphogenetic protein 15 both activate development of human primordial follicles in vitro, with seemingly more beneficial effects of GDF9. J Clin Endocrinol Metab. 2011;96(8):E1246–54. Scholar
  27. 27.
    Feuerstein P, Cadoret V, Dalbies-Tran R, Guerif F, Bidault R, Royere D. Gene expression in human cumulus cells: one approach to oocyte competence. Hum Reprod. 2007;22(12):3069–77. Scholar
  28. 28.
    McKenzie LJ, Pangas SA, Carson SA, Kovanci E, Cisneros P, Buster JE, et al. Human cumulus granulosa cell gene expression: a predictor of fertilization and embryo selection in women undergoing IVF. Hum Reprod. 2004;19(12):2869–74. Scholar
  29. 29.
    Cillo F, Brevini TA, Antonini S, Paffoni A, Ragni G, Gandolfi F. Association between human oocyte developmental competence and expression levels of some cumulus genes. Reproduction. 2007;134(5):645–50. Scholar
  30. 30.
    Anderson RA, Sciorio R, Kinnell H, Bayne RA, Thong KJ, de Sousa PA, et al. Cumulus gene expression as a predictor of human oocyte fertilisation, embryo development and competence to establish a pregnancy. Reproduction. 2009;138(4):629–37. Scholar
  31. 31.
    Feuerstein P, Puard V, Chevalier C, Teusan R, Cadoret V, Guerif F, et al. Genomic assessment of human cumulus cell marker genes as predictors of oocyte developmental competence: impact of various experimental factors. PLoS One. 2012;7(7):e40449. Scholar
  32. 32.
    Kordus RJ, LaVoie HA. Granulosa cell biomarkers to predict pregnancy in ART: pieces to solve the puzzle. Reproduction. 2017;153(2):R69–83. Scholar
  33. 33.
    Wathlet S, Adriaenssens T, Segers I, Verheyen G, Janssens R, Coucke W, et al. New candidate genes to predict pregnancy outcome in single embryo transfer cycles when using cumulus cell gene expression. Fertil Steril. 2012;98(2):432–9 e1–4. Scholar
  34. 34.
    Burnik Papler T, Vrtacnik Bokal E, Maver A, Lovrecic L. Specific gene expression differences in cumulus cells as potential biomarkers of pregnancy. Reprod BioMed Online. 2015;30(4):426–33. Scholar
  35. 35.
    Sanchez F, Smitz J. Molecular control of oogenesis. Biochim Biophys Acta. 2012;1822(12):1896–912. Scholar
  36. 36.
    Wissing ML, Kristensen SG, Andersen CY, Mikkelsen AL, Host T, Borup R, et al. Identification of new ovulation-related genes in humans by comparing the transcriptome of granulosa cells before and after ovulation triggering in the same controlled ovarian stimulation cycle. Hum Reprod. 2014;29(5):997–1010. Scholar
  37. 37.
    Luo CW, Kawamura K, Klein C, Hsueh AJ. Paracrine regulation of ovarian granulosa cell differentiation by stanniocalcin (STC) 1: mediation through specific STC1 receptors. Mol Endocrinol. 2004;18(8):2085–96. Scholar
  38. 38.
    Deol HK, Varghese R, Wagner GF, Dimattia GE. Dynamic regulation of mouse ovarian stanniocalcin expression during gestation and lactation. Endocrinology. 2000;141(9):3412–21. Scholar
  39. 39.
    Vaiarelli A, Cimadomo D, Patrizio P, Venturella R, Orlando G, Soscia D, et al. Biochemical pregnancy loss after frozen embryo transfer seems independent of embryo developmental stage and chromosomal status. Reprod BioMed Online. 2018;37(3):349–57. Scholar
  40. 40.
    Rienzi L, Ubaldi F, Anniballo R, Cerulo G, Greco E. Preincubation of human oocytes may improve fertilization and embryo quality after intracytoplasmic sperm injection. Hum Reprod. 1998;13(4):1014–9.Google Scholar
  41. 41.
    Ubaldi F, Anniballo R, Romano S, Baroni E, Albricci L, Colamaria S, et al. 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):1199–205. Scholar
  42. 42.
    Capalbo A, Rienzi L, Cimadomo D, Maggiulli R, Elliott T, Wright G, et al. Correlation between standard blastocyst morphology, euploidy and implantation: an observational study in two centers involving 956 screened blastocysts. Hum Reprod. 2014;29(6):1173–81. Scholar
  43. 43.
    Capalbo A, Treff NR, Cimadomo D, Tao X, Upham K, Ubaldi FM, et al. Comparison of array comparative genomic hybridization and quantitative real-time PCR-based aneuploidy screening of blastocyst biopsies. Eur J Hum Genet. 2015;23(7):901–6. Scholar
  44. 44.
    Treff NR, Tao X, Ferry KM, Su J, Taylor D, Scott RT Jr. Development and validation of an accurate quantitative real-time polymerase chain reaction-based assay for human blastocyst comprehensive chromosomal aneuploidy screening. Fertil Steril. 2012;97(4):819–24. Scholar
  45. 45.
    Gardner DK, Schoolcraft B. In vitro culture of human blastocyst. In: Jansen R, Mortimer D, editors. Towards reproductive certainty: infertility and genetics beyond. Carnforth: Parthenon Press; 1999. p. 377–88.Google Scholar
  46. 46.
    Cimadomo D, Capalbo A, Levi-Setti PE, Soscia D, Orlando G, Albani E, et al. Associations of blastocyst features, trophectoderm biopsy and other laboratory practice with post-warming behavior and implantation. Hum Reprod. 2018;33:1992–2001. Scholar
  47. 47.
    Braude P, Bolton V, Moore S. Human gene expression first occurs between the four- and eight-cell stages of preimplantation development. Nature. 1988;332(6163):459–61. Scholar
  48. 48.
    Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002;3(7):RESEARCH0034.Google Scholar
  49. 49.
    Wathlet S, Adriaenssens T, Segers I, Verheyen G, Van de Velde H, Coucke W, et al. Cumulus cell gene expression predicts better cleavage-stage embryo or blastocyst development and pregnancy for ICSI patients. Hum Reprod. 2011;26(5):1035–51. Scholar
  50. 50.
    Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 2008;3(6):1101–8.Google Scholar
  51. 51.
    McCall MN, McMurray HR, Land H, Almudevar A. On non-detects in qPCR data. Bioinformatics. 2014;30(16):2310–6. Scholar
  52. 52.
    Adriaenssens T, Wathlet S, Segers I, Verheyen G, De Vos A, Van der Elst J, et al. Cumulus cell gene expression is associated with oocyte developmental quality and influenced by patient and treatment characteristics. Hum Reprod. 2010;25(5):1259–70. Scholar
  53. 53.
    Green KA, Franasiak JM, Werner MD, Tao X, Landis JN, Scott RT Jr, et al. Cumulus cell transcriptome profiling is not predictive of live birth after in vitro fertilization: a paired analysis of euploid sibling blastocysts. Fertil Steril. 2018;109(3):460–6 e2. Scholar
  54. 54.
    Adriaenssens T, Segers I, Wathlet S, Smitz J. The cumulus cell gene expression profile of oocytes with different nuclear maturity and potential for blastocyst formation. J Assist Reprod Genet. 2011;28(1):31–40. Scholar
  55. 55.
    Park MH, Nishimura K, Zanelli CF, Valentini SR. Functional significance of eIF5A and its hypusine modification in eukaryotes. Amino Acids. 2010;38(2):491–500. Scholar
  56. 56.
    Fujimura K, Choi S, Wyse M, Strnadel J, Wright T, Klemke R. Eukaryotic translation initiation factor 5A (EIF5A) regulates pancreatic cancer metastasis by modulating RhoA and Rho-associated kinase (ROCK) protein expression levels. J Biol Chem. 2015;290(50):29907–19. Scholar
  57. 57.
    Qin X, Liang Y, Guo Y, Liu X, Zeng W, Wu F, et al. Eukaryotic initiation factor 5A and Ca(2+) /calmodulin-dependent protein kinase 1D modulate trophoblast cell function. Am J Reprod Immunol. 2018;80(1):e12845. Scholar
  58. 58.
    Haney S, Zhao J, Tiwari S, Eng K, Guey LT, Tien E. RNAi screening in primary human hepatocytes of genes implicated in genome-wide association studies for roles in type 2 diabetes identifies roles for CAMK1D and CDKAL1, among others, in hepatic glucose regulation. PLoS One. 2013;8(6):e64946. Scholar
  59. 59.
    Bergamaschi A, Kim YH, Kwei KA, La Choi Y, Bocanegra M, Langerod A, et al. CAMK1D amplification implicated in epithelial-mesenchymal transition in basal-like breast cancer. Mol Oncol. 2008;2(4):327–39. Scholar
  60. 60.
    Eppig JJ. Prostaglandin E2 stimulates cumulus expansion and hyaluronic acid synthesis by cumuli oophori isolated from mice. Biol Reprod. 1981;25(1):191–5.Google Scholar
  61. 61.
    Hizaki H, Segi E, Sugimoto Y, Hirose M, Saji T, Ushikubi F, et al. Abortive expansion of the cumulus and impaired fertility in mice lacking the prostaglandin E receptor subtype EP(2). Proc Natl Acad Sci U S A. 1999;96(18):10501–6.Google Scholar
  62. 62.
    Lim H, Paria BC, Das SK, Dinchuk JE, Langenbach R, Trzaskos JM, et al. Multiple female reproductive failures in cyclooxygenase 2-deficient mice. Cell. 1997;91(2):197–208.Google Scholar
  63. 63.
    Davis BJ, Lennard DE, Lee CA, Tiano HF, Morham SG, Wetsel WC, et al. Anovulation in cyclooxygenase-2-deficient mice is restored by prostaglandin E2 and interleukin-1beta. Endocrinology. 1999;140(6):2685–95. Scholar
  64. 64.
    Takahashi T, Morrow JD, Wang H, Dey SK. Cyclooxygenase-2-derived prostaglandin E(2) directs oocyte maturation by differentially influencing multiple signaling pathways. J Biol Chem. 2006;281(48):37117–29. Scholar
  65. 65.
    Fulop C, Salustri A, Hascall VC. Coding sequence of a hyaluronan synthase homologue expressed during expansion of the mouse cumulus-oocyte complex. Arch Biochem Biophys. 1997;337(2):261–6. Scholar
  66. 66.
    Salustri A, Yanagishita M, Hascall VC. Synthesis and accumulation of hyaluronic acid and proteoglycans in the mouse cumulus cell-oocyte complex during follicle-stimulating hormone-induced mucification. J Biol Chem. 1989;264(23):13840–7.Google Scholar
  67. 67.
    Salustri A, Ulisse S, Yanagishita M, Hascall VC. Hyaluronic acid synthesis by mural granulosa cells and cumulus cells in vitro is selectively stimulated by a factor produced by oocytes and by transforming growth factor-beta. J Biol Chem. 1990;265(32):19517–23.Google Scholar
  68. 68.
    Alfarawati S, Fragouli E, Colls P, Stevens J, Gutierrez-Mateo C, Schoolcraft WB, et al. The relationship between blastocyst morphology, chromosomal abnormality, and embryo gender. Fertil Steril. 2011;95(2):520–4. Scholar
  69. 69.
    Fragouli E, Wells D, Iager AE, Kayisli UA, Patrizio P. Alteration of gene expression in human cumulus cells as a potential indicator of oocyte aneuploidy. Hum Reprod. 2012;27(8):2559–68. Scholar
  70. 70.
    Fragouli E, Lalioti MD, Wells D. The transcriptome of follicular cells: biological insights and clinical implications for the treatment of infertility. Hum Reprod Update. 2014;20(1):1–11. Scholar
  71. 71.
    McCoy RC, Demko Z, Ryan A, Banjevic M, Hill M, Sigurjonsson S, et al. Common variants spanning PLK4 are associated with mitotic-origin aneuploidy in human embryos. Science. 2015;348(6231):235–8. Scholar
  72. 72.
    McCoy RC, Demko ZP, Ryan A, Banjevic M, Hill M, Sigurjonsson S, et al. Evidence of selection against complex mitotic-origin aneuploidy during preimplantation development. PLoS Genet. 2015;11(10):e1005601. Scholar
  73. 73.
    McCoy RC, Newnham LJ, Ottolini CS, Hoffmann ER, Chatzimeletiou K, Cornejo OE, et al. Tripolar chromosome segregation drives the association between maternal genotype at variants spanning PLK4 and aneuploidy in human preimplantation embryos. Hum Mol Genet. 2018;27(14):2573–85. Scholar
  74. 74.
    Basile N, Vime P, Florensa M, Aparicio Ruiz B, Garcia Velasco JA, Remohi J, et al. The use of morphokinetics as a predictor of implantation: a multicentric study to define and validate an algorithm for embryo selection. Hum Reprod. 2015;30(2):276–83. Scholar
  75. 75.
    Betts DH, Madan P. Permanent embryo arrest: molecular and cellular concepts. Mol Hum Reprod. 2008;14(8):445–53. Scholar
  76. 76.
    Hammond ER, Stewart B, Peek JC, Shelling AN, Cree LM. Assessing embryo quality by combining non-invasive markers: early time-lapse parameters reflect gene expression in associated cumulus cells. Hum Reprod. 2015;30(8):1850–60. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.DAHFMO, Unit of Histology and Medical EmbryologySapienza, University of RomeRomeItaly
  2. 2.Casa di cura Villa SalariaRomeItaly
  3. 3.Clinica Valle Giulia, G.EN.E.R.A. Centers for Reproductive MedicineRomeItaly
  4. 4.IgenomixMarosticaItaly

Personalised recommendations