Trophectoderm biopsy protocols can affect clinical outcomes: time to focus on the blastocyst biopsy technique

Abstract Study question Do different trophectoderm biopsy techniques affect mosaicism rate in human blastocysts? Summary answer No statistical significance was found between biopsy techniques and mosaicism rate. However, an increase in mosaicism rate was observed when the flicking technique was used. What is known already Mosaicism is defined as two or more distinct cell lines within an embryo. Recent advances in Next Generation Sequencing (NGS) technology with an increased sensitivity allows a higher accuracy in quantification of mosaic levels in biopsied cells. The incidence of mosaicism is widely debated as there are many attributing technical and biological factors. Since, trophectoderm biopsy is a technically challenging process, it is crucial to ensure that the both biopsied cells and blastocyst suffers minimal damage during biopsy. Study design, size, duration This is a prospective study involving 222 patients (age range= 18–44, mean age= 31.5) who underwent IVF cycles in Alpha IVF, Malaysia from March 2019 to August 2019. Six hundred and sixty-eight (668) of the blastocysts were biopsied on Day 5 (Group 1) while 177 blastocysts were biopsied on Day 6 (Group 2). The blastocysts in these groups were further categorised into their corresponding biopsy techniques: (A) laser+pulling; (B) laser+flicking; (C) flicking only. Participants/materials, setting, methods Blastocysts which were at least fair graded (Gardner, 1999) were biopsied and vitrified (Cryotec, Japan). The number of biopsied cells ranged from 5 to 10 cells. All biopsied trophectoderm samples were subjected to Preimplantation Genetic Testing for Aneuploidy (PGT-A) with Next Generation Sequencing (NGS) (Ion Torrent, USA). Chromosomal mosaicism analysis was done using ReproSeq Mosaic PGS w1.1 workflow. Trophectoderm biopsied sample which were tested to have 20% to 80% aneuploid cells were reported as mosaic. Main results and the role of chance In Group 1, the mosaicism rates for biopsy technique A, B and C were 23.3% (104/446), 28.2% (58/206) and 37.5% (6/16) respectively. In Group 2, the mosaicism rates for biopsy technique A, B and C were 14.6% (7/48), 19.5% (23/118) and 27.3% (3/11) respectively. There were no significant differences (p > 0.05) in mosaicism rates between all study groups and subgroups. Limitations, reasons for caution Although no statistical significance was found between trophectoderm biopsy techniques and the prevalence of mosaicism, there is a trend of an increase in mosaicism rate when the flicking technique was used. Therefore, further studies with a larger sample size should be undertaken. Wider implications of the findings: Our study demonstrates a trend in the decrease of mosaicism rate when laser pulses was used to loosen the cell junction of targeted cells. Hence, in place of the flicking method alone, laser pulses should be applied during trophectoderm biopsy if our findings are confirmed in a larger controlled study. Trial registration number Not applicable

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Dual trophectoderm biopsy on the same blastocyst does not impair clinical outcomes

Fertility and Sterility ◽

10.1016/j.fertnstert.2015.07.577 ◽

2015 ◽

Vol 104 (3) ◽

pp. e186 ◽

Cited By ~ 2

Author(s):

J.E. Swain ◽

W.B. Schoolcraft ◽

M. Katz-Jaffe

Keyword(s):

Clinical Outcomes ◽

Trophectoderm Biopsy

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Prolonged interval time between blastocyst biopsy and vitrification compromised the outcomes in preimplantation genetic testing

Zygote ◽

10.1017/s0967199420000866 ◽

2021 ◽

pp. 1-6

Author(s):

Shun Xiong ◽

Jun Xia Liu ◽

Dong Yun Liu ◽

Jia Hong Zhu ◽

Xiang Wei Hao ◽

...

Keyword(s):

Genetic Testing ◽

Clinical Outcomes ◽

Pregnancy Rates ◽

Clinical Pregnancy ◽

Ongoing Pregnancy ◽

Miscarriage Rate ◽

Preimplantation Genetic Testing ◽

Embryo Survival ◽

Blastocyst Biopsy ◽

Group 1

Summary This study aimed to evaluate to what extent the different interval times between trophectoderm (TE) biopsy and vitrification influence the clinical outcomes in preimplantation genetic testing (PGT) cycles. Patients who underwent frozen embryo transfer (FET) after PGT between 2015 and 2019 were recruited. In total, 297 cycles with single day 5 euploid blastocyst transfer were included. These cycles were divided into three groups according to the interval times: <1 h group, 1–2 h group, and ≥2 h group. Blastocyst survival, clinical pregnancy, miscarriage, and ongoing pregnancy rates were compared. The results showed that, in PGT-SR cycles, survival rate in the ≥2 h group (96.72%) was significantly lower than in the <1 h group (100%, P = 0.047). The clinical pregnancy rate in the ≥2 h group was 55.93%, significantly lower than in the <1 h group (74.26%, P = 0.017). The ongoing pregnancy rates in the 1–2 h group and the ≥2 h group were 48.28% and 47.46%, respectively, significantly lower than that in the <1 h group (67.33%, P < 0.05). The miscarriage rate in the 1–2 h group was 18.42%, significantly higher than that in the <1 h group (5.33%, P = 0.027). In PGT-A cycles, the clinical pregnancy and ongoing pregnancy rates in the <1 h group were 67.44% and 53.49%, respectively, higher than that in the 1–2 h group (52.94%, 47.06%, P > 0.05) and the ≥2 h group (52.63%, 36.84%, P > 0.05). In conclusion, vitrification of blastocysts beyond 1 h after biopsy significantly influences embryo survival and clinical outcomes and is therefore not recommended.

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A novel trophectoderm biopsy technique for all blastocyst stages

Reproductive Medicine and Biology ◽

10.1002/rmb2.12418 ◽

2021 ◽

Author(s):

Yamato Mizobe ◽

Yukari Kuwatsuru ◽

Yuko Kuroki ◽

Yumiko Fukumoto ◽

Mari Tokudome ◽

...

Keyword(s):

Trophectoderm Biopsy ◽

Biopsy Technique

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Can the location of a trophectoderm biopsy contribute to human blastocyst development ?

10.1101/109298 ◽

2017 ◽

Author(s):

Tomoe Takano ◽

Miyako Funabiki ◽

Sagiri Taguchi ◽

Fumie Saji ◽

Namiko Amano ◽

...

Keyword(s):

Clinical Outcomes ◽

Clinical Evidence ◽

Inner Cell Mass ◽

Cell Mass ◽

Multivariate Logistic Regression Analysis ◽

Blastocyst Transfer ◽

Trophectoderm Biopsy ◽

Blastocyst Development ◽

Human Blastocyst ◽

Single Blastocyst Transfer

AbstractThe influence of the location of a trophectoderm biopsy in human blastocysts on the development of those blastocysts has not yet been investigated. In our prospective study (n=92), our multivariate logistic regression analysis indicated that blastocoel development was influenced by the location of the trophectoderm biopsy (p=0.049) and by the type of human blastocyst used (fresh or thawed) (p=0.037), regardless of the patient’s age (p=0.507) and the number of days for the human blastocyst in the pretrophectoderm biopsy (p=0.239). Therefore, when a trophectoderm biopsy is close to the inner cell mass (ICM) in human blastocysts, it improves the progress of blastocoel development.Clinical evidence suggests that the progress of blastocoel development is a predictor of clinical outcomes after single blastocyst transfer. Therefore, when the trophectoderm biopsy is done from near the ICM, improvement of clinical outcomes after single blastocyst transfer may be expected.

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