Introduction: Preimplantation Genetic Testing is one of the methods to screen genetic defects in embryos created through In Vitro Fertilization. In developing country this technique is still new and expensive method. Thus, knowing several factors in predicting the occurrence of three-pronuclear zygote can help much.Method: 472 cycles in 449 subjects who underwent controlled ovarian hyperstimulation in IVF cycles between January 2013 and August 2014 were included in the study. Categorical variables were compared using Chi Square test and continuous variables were analyzed using Independent t-test, and statistically significant was considered when p &lt; 0.05. Multivariate logistic regression analysis was performed in order to correlate clinical variables and the occurrence of three-pronuclear zygote (3pn). Results: There were 38 3pn was identified in this study (8.05%). No correlation was found between age of the women, number of mature follicles, which are less than six with the incidence of 3pn. However, level of Anti Mullerian Hormones (AMH) found to be the strongest predictor with the incidence of 3PN (p &lt; 0,01, RR 2.5, CI95% 1,55; 4.16).Conclusion: level of AMH is known to be strong predictor of thee pronuclear zygote after IVF cycle

Agnes Susanto, Cynthia

Wiweko, Budi

English

KnE Publishing Platform

 KnE MedicineASPIRE Conference ProceedingsThe 6th Congress of the Asia Pacific Initiative onReproduction (ASPIRE 2016), Volume 2016Conference PaperPredictive Factors for Three PronuclearZygote in IVF CycleCynthia Agnes Susanto and Budi WiwekoFaculty of Medicine, University of Indonesia, Obstetrics and Gynecology Department, Jakarta,IndonesiaAbstractIntroduction. Preimplantation Genetic Testing is one of the methods to screen geneticdefects in embryos created through In Vitro Fertilization. In developing countrythis technique is still new and expensive method. Thus, knowing several factorsin predicting the occurrence of three-pronuclear zygote can help much. Method.472 cycles in 449 subjects who underwent controlled ovarian hyperstimulationin IVF cycles between January 2013 and August 2014 were included in the study.Categorical variables were compared using Chi Square test and continuous variableswere analyzed using Independent t-test, and statistically significant was consideredwhen p < 0.05. Multivariate logistic regression analysis was performed in orderto correlate clinical variables and the occurrence of three-pronuclear zygote (3pn).Results. There were 38 3pn was identified in this study (8.05%). No correlation wasfound between age of the women, number of mature follicles, which are less than sixwith the incidence of 3pn. However, level of Anti Mullerian Hormones (AMH) foundto be the strongest predictor with the incidence of 3PN (p < 0,01, RR 2.5, CI95% 1,55;4.16). Conclusion. level of AMH is known to be strong predictor of thee pronuclearzygote after IVF cycle.Keywords: Preimplantation genetic testing, IVF, three pronuclear, zygote, Anti-Mullerian Hormones1. IntroductionA common definition of infertility has been used in clinical settings is: “One year ofunwanted non-conception with unprotected intercourse in the fertile phase of themenstrual cycles [1].”Based onWHO, there are around 186millionmarried womanwith infertility in devel-oping country who is either having primary infertility or secondary infertility [2]. Whilein Indonesia, Yasmin Clinic that is located in Cipto Mangunkusumo General Hospital,Jakartam showing increasing of couple visit to get management of infertility from 1238couples in 20013 to 1834 couples in 2014.In Vitro Fertilization (IVF) becomes one of alternative method used in order to con-ceived. Pregnancy rate in Yasmin clinic Kencana is around 40 – 45% from all of thecouple underwent IVF. In order to undergo IVF cycles, several factors contributes tothe successfulness of it, for example the age of the patient, quality of endometrium,quality of embryos. Several factors such as age of the women, concentration of sperm,How to cite this article: Cynthia Agnes Susanto and Budi Wiweko, (2016) “Predictive Factors for Three Pronuclear Zygote in IVF Cycle,” in ASPIREConference Proceedings, The 6th Congress of the Asia Pacific Initiative on Reproduction, KnE Medicine, pages 12–17. DOI 10.18502/kme.v1i1.531 Page 12Corresponding Author: BudiWiweko; email:wiwekobudi@yahoo.co.idReceived: 24 August 2016Accepted: 25 September 2016Published: 4 October 2016Publishing services providedby Knowledge ECynthia Agnes Susantoand Budi Wiweko. This articleis distributed under the termsof the Creative CommonsAttribution License, whichpermits unrestricted use andredistribution provided thatthe original author andsource are credited.Selection and Peer-reviewunder the responsibility ofthe ASPIRE ConferenceCommittee. KnE Medicine ASPIRE Conference Proceedingshistory of recurrent pregnancy loss, and prior failure of IVF known to effect the qualitiesof embryos. Objectively, quality of embryo can be evaluated by the use of pre implan-tation screening. This type of screening is used in order to detect genetic materials ormorphological examinations [3]. To date, in Indonesia, qualities of embryo are checkedbased onmorphologic appearance other than genetic screening. Morphological testingis still the chosen option to choose quality of embryos in short period of time. Whilepreimplantation genetic screening is not being used routinely in Indonesia since it isexpensive and couple needs to pay extra.Previous studies shows pronucleus cells is one of the good predictor of evaluatingzygote development. Around 5% from zygote came from In Vitro Fertilization methodhas more than two pronucleus (with highest rank is three pronucleus (3pn)). Thisabnormality is happened due to more than one penetration of sperm, or failure ofextrusion from second polar body. The most common cause of this abnormalities isthe penetration of more than two sperm into a single ovum, which causing embryosto have triploid chromosome of XXX, XXY, or XYY. This type of zygote will not undergoembryo transfer in IVF. The prevalence of 3PN is one to three percent of all embryos.Abnormalities are happening within the early stage of development, though some ofthe abnormalities might developed in later days [4].Several examinations previously mention has purpose to evaluate anomalies withinembryos before it get transferred inside a womb. However, there is no known riskfactors that can cause conditions of 3pn zygote. Thus, knowing several factors in pre-dicting the occurrence of three-pronuclear zygote can help much.2. MethodThe subjects of the study were 472 cycles in 449 subjects who underwent controlledovarian hyperstimulation in IVF cycles at Yasmin Clinic of Dr. Cipto MangunkusumoGeneral Hospital, Jakarta, Indonesia between January 2013 and August 2014. The exclu-sion criteria were patients who did not follow the protocol of IVF to the completion andwhich the data is incomplete.Data of medical records were recorded in the study forms which have been madespecifically to facilitate data entry and analysis. The data were processed using IBMSPSS for windows version 20.At this stage of analysis, we studied the relationship between two variables com-prising the comparison between clinical variables, such as patient’s age, type of infer-tility, causes of infertility in female and male, basal AMH level, and number of matureoocyte. Categorical variables were compared using Chi Square test and continuousvariables were analyzed using Independent t-test, and statistically significant wasconsidered when p < 0.05.DOI 10.18502/kme.v1i1.531 Page 13 KnE Medicine ASPIRE Conference ProceedingsCharacteristics N (%)Age 20-25 12 (2.5)26-30 76 (16.1)31-35 136 (28.8)36-40 164 (34.7)>40 84 (17.8)Causes of Infertility Female 437 (92.6)Male 35 (7.4)AMH level ≤ 1.4 98 (20.8)>1.4 374 (79.2)Number of Mature follicles ≤6 133 (28.2)>6 339 (71.8)Female factor Endometriosis 61 (13.9)PCOS 56 (12.9)Tubal factor 69 (15.7)DOR/PoorResponder137 (31.1)Hyperprolactinemia7 (1.6)Others 110 (25)T 1: Demographic data between IVF cycles (total of 472 cycles).3. ResultsThere were four hundred seventy two cycles of IVF were performed in 449 couplesundergoing IVF program from January 2013 and August 2014. The distribution ofpatients’ ages during period of IVF program was between the ages of 20 until 48.Mean age of the patients was 35.5 years of age with mainly cause of infertility isfemale with poor responders.Based on this study, it was found total of 38 cycles (8.05%) found to have 3pnembryos. Risk factors such as patient’s age, number of mature oocyte and level orAMH were compared using Chi Square test (Table 2).Based on Table 2, statistically significant findings to the incidence of 3pn is onlylevel of AMH below 1,4. Figure 3 describes mean of each categorical dependent factor.Mean of age of patient whose having 3pn zygote is 36.49 compared to normal zygoteof 35.40. Also number of mature follicle found to be more in 3pn zygotes with meanof 14.87. However, level of AMH in-patient present with 3pn zygote found to be lowercompared to normal zygotes.On Table 3, it is shown relative risk of level of AMH to the incidence of 3pn is 2.5 (CI95% 1.56; 4.16).DOI 10.18502/kme.v1i1.531 Page 14 KnE Medicine ASPIRE Conference ProceedingsParameter p-valueAge 0.08Number of maturefolicles0.133Level of AMH (< 1.4) < 0.001T 2: Value of p of risk factors influence the development of 3pn zygote.Figure 1: Mean value of each risk factors in normal group and 3pn group4. DiscussionThree pronuclear can happen when there is a fertilization of more than one sperm orthere is failure meiosis of an oocyte. The cause of 3pn until today is still debatable.Some expert believe 3pn can be cause by maternal age or sperm abnormalities [5-7].In this study, there were 472 cycles of IVF from 449 couple undergoing IVF cycles.There is 8.05 % of three pronuclear zygote can be found in this study. This numberseems more than usual prevalence of 3pn worldwide. This study found there is nosignificant correlation between the incidence of 3pn with maternal age (p=0.08) andnumber of mature follicle (p = 0.133). This result is consistent with previous study by Liand friends in 2015, which found there is no significant correlation between maternalages with the incidence of 3pn [5].There is a statistically significant finding between levels of AMH with the incidenceof 3pn zygotes (p < 0.01). In women, AMH is a hormone works in the cell trough tworeceptors that is AMH receptor type I and tipe II (AMHRI and AMHRII). AMH has twomain function in female reproductive system, which is within primary recruitment,AMH level 3PN Zygote+ -<1,4 22 76>1,4 33 341T 3: Two by two table showing relationship of level of AMH with incidence of 3pn zygote.DOI 10.18502/kme.v1i1.531 Page 15 KnE Medicine ASPIRE Conference ProceedingsAMH will inhibit primordial follicle recruitment into the growing pool and during cyclicrecruitment, AMH will decrease the sensitivity of ovarian follicle to Follicle StimulatingHormone (FSH), thus AMH known to be one of the factor which regulates the growingof the follicle, thus it will influence the oocyte qualities which then will influence thesuccessfulness of IVF [8,9].To date, there is no single study which trying to find the relationship between levelof AMH with the incidence of 3pn zygote. However, level of AMH is proven to be useas an indicator to evaluate oocyte viability in IVF and low level of AMH can predict theoccurrence of aneuploidy [10,11].5. ConclusionAs a conclusion, level of AMH is known to be strong predictor of thee pronuclear zygoteafter IVF cycle. However, this preliminary study can be useful to conduct further studiesin order to evaluate more risk factors of three-pronuclear zygote.References[1] A. E. Puscheck, Infertility [article on internet] 2015 [cited 3 March 2016]. Availablefrom: http://emedicine.medscape.com/article/-overview.[2] S. O. Rutstein, O. R. Marco, and I. H. Shah, Infenduncity, infertility, and childlessnessin developing countries, World Health Organization: Calverton, 2004.[3] A. Nicoli, F. Capodanno, L. Moscato, I. Rondini, M. T. Villani, A. Tuzio, and G. B.La Sala, Analysis of pronuclear zygote configurations in 459 clinical pregnanciesobtainedwith assisted reproductive technique procedures, Reproductive Biology andEndocrinology, 8, article no. 77, (2010).[4] C. Staessen and A. C. Van Steirteghem, The chromosomal constitution of embryosdeveloping from abnormally fertilized oocytes after intracytoplasmic sperminjection and conventional in-vitro fertilization, Human Reproduction, 12, no. 2, 321–327, (1997).[5] M. Li, W. Zhao, X. Xue, S. Zhang, W. Shi, and J. Shi, Three pro-nuclei (3PN) incidencefactors and clinical outcomes: A retrospective study from the fresh embryo transferof in vitro fertilization with donor sperm (IVF-D), International Journal of Clinical andExperimental Medicine, 8, no. 8, 13997–14003, (2015).[6] B. E. Rosenbusch, Mechanisms giving rise to triploid zygotes during assistedreproduction, Fertility and Sterility, 90, no. 1, 49–55, (2008).[7] D. E.McFadden andW. P. Robinson, Phenotype of triploid embryos, Journal of MedicalGenetics, 43, no. 7, 609–612, (2006).[8] M. J. G. Gruijters, J. A. Visser, A. L. L. Durlinger, and A. P. N. Themmen, Anti-Müllerianhormone and its role in ovarian function, Molecular and Cellular Endocrinology, 211,no. 1-2, 85–90, (2003).[9] A. L. L. Durlinger, J. A. Visser, and A. P. N. Themmen, Regulation of ovarian function:The role of anti-Müllerian hormone, Reproduction, 124, no. 5, 601–609, (2002).DOI 10.18502/kme.v1i1.531 Page 16 KnE Medicine ASPIRE Conference Proceedings[10] B. N. Mehta, M. N. Chimote, N. N. Chimote, N. M. Nath, and N. M. Chimote,Follicular-fluid anti-Mullerian hormone (FF AMH) is a plausible biochemical indicatorof functional viability of oocyte in conventional in vitro fertilization (IVF) cycles,Journal of Human Reproductive Sciences, 6, no. 2, 99–105, (2013).[11] AP. Melnick, EM. Murphy, H. Liu, OK. Davis, and Z. Rosenwaks, Low AMH isassociated with increased aneuploidy in women 40 and younger. Fertil steril, inRosenwaks Z. Low AMH is associated with increased aneuploidy in women 40 andyounger. Fertil steril, 104–248, 104, 248-249, 2015.DOI 10.18502/kme.v1i1.531 Page 17

Predictive Factors For Three Pronuclear Zygote In Ivf Cycle

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Predictive Factors For Three Pronuclear Zygote In Ivf Cycle

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