Study of Morphokinetics in Day 3 Embryo with Implantation Potential and Effect of Sperm Cryopreservation on Embryogenesis

Aim: In recent past, many studies had come up with the combination of time-lapse (TL) imaging of embryo morphokinetics as a noninvasive means for improving embryo selection and in vitro fertilization (IVF) success. The primary objective of the study was to find out if there is significant variation in morphokinetics of embryos with different implantation potential and also to study the effect of sperm freezing on time points of embryogenesis events in embryos with implantation potential. Materials and methods: Kinetic data and cycle outcomes were analyzed retrospectively in 142 patients who had undergone IVF/intracytoplasmic sperm injection (ICSI) cycles using semen with normal parameters and embryo transfer (ET) on day 3. For the surety of specificity of morphokinetics, only cases with single ET cycles were included in the study. Timing of specific events, from the point of ICSI, was determined using TL imaging. Kinetic markers like time to syngamy (t-pnf), t2, time to two cells (c), 3c (t3), 4c (t4), 5c (t5), 8c (t8), tMor, CC2, CC3, t5–t2, t5–t4, s1, s2, and s3 were calculated. The cleavage synchronicity from the 2–8 cell stage (CS2–8), from 4 to 8 cell stage (CS4–8), and from 2 to 4 cell stage (CS2–4) were calculated as defined elsewhere. Deoxyribonucleic acid replication time ratio (DR) was also included RESEARCH ARTiCLE 1Lab Director, 2Gynecologist and Medical Director, 3Senior Embryologist, 4Research Scientist, 5,7Gynecologist, 6Gynecologist and Director, 8Statistician, 9Assistant Professor, 10Former Assistant Professor and Head 1Department of IVF, Akanksha Hospital & Research Institute Anand, Gujarat, India; Department of Biochemistry, P D Patel Institute of Applied Sciences, Changa, Gujarat, India 2,5-7Department of Obstetrics and Gynecology, Akanksha Hospital & Research Institute, Anand, Gujarat, India 3Department of IVF, Akanksha Hospital & Research Institute Anand, Gujarat, India 4Department of Research and Development, Sat Kaival Hospital Private Limited, Anand, Gujarat, India 8Department of Administration, Sardar Patel University, Anand Gujarat, India 9Department of Biotechnology, P D Patel Institute of Applied Sciences, Changa, Gujarat, India 10Department of Biochemistry, P D Patel Institute of Applied Sciences, Changa, Gujarat, India Corresponding Author: Harsha K Bhadarka, Lab Director Department of IVF, Akanksha Hospital & Research Institute Anand, Gujarat, India; Department of Biochemistry, PD Patel Institute of Applied Sciences, Changa, Gujarat, India, Phone: +919727416492, e-mail: harshabhadarka@yahoo.co.in 10.5005/jp-journals-10016-1150 in the comparison. Analysis of variance test was used for comparison of the mean timing of cell division and cell cycle intervals. Results: Morphokinetics t-pnf, t2, t8, CC2, S2, S3, CS2–8, CS4–8, and CS2–4 differed significantly between embryos with and without implantation potential, when embryos were developed using fresh semen, while t3, t4, t5, CC2, S2, t5–t2, CS2–4, and DR differed significantly between the embryos with and without implantation potential when frozen semen was used. No significant difference was found in mean value of any of the above-stated parameters when comparison was done between implanted embryos fertilized by either fresh or cryopreserved sperm. Conclusion: Many morphokinetics parameters of embryogenesis vary significantly between embryos with different ability to implant; therefore, the criteria developed in our IVF lab can be useful for selection of suitable embryo even at day 3 of development with more chances of implantation. Clinical significance: Study indicates necessity of development of individualized selection model based on morphokinetics for every IVF lab and also confirms freezing as an important tool for fertility preservation of males as it does not affect events of embryogenesis.