OBJECTIVE: Does artificial shrinkage before fresh blastocyst transfer improve clinical pregnancy rates in IVF?
METHODS: In this monocentric prospective, randomized, double-blind, controlled pilot study, 150 couples undergoing fresh single-blastocyst transfer were randomized between 20 May 2018 and 22 February 2022. In the artificial shrinkage group (AS group), a single laser pulse was directed to the cellular junction of the trophectoderm on the opposite side of the inner cell mass in each blastocyst. IVF outcomes were clinical pregnancy, multiple pregnancy and live birth rates. Cell-free DNA (cfDNA) concentration was also measured by quantitative real-time PCR in the blastocyst culture medium.
RESULTS: In total, 142 couples underwent fresh single-blastocyst transfer: control group, no artificial shrinkage, n = 47; and AS group, artificial shrinkage, n = 95; An intention-to-treat (ITT) analysis was employed. After a reassessment and the exclusion of patients with major protocol deviations, 139 couples underwent fresh single-blastocyst transfer under optimal conditions: control group, n = 47; and AS group, n = 92; a per-protocol analysis was used here. The clinical and laboratory characteristics were not significantly different between the groups. The clinical pregnancy rate was similar in the control and AS groups (ITT: 48.9% versus 49.5%, P = 0.97; per protocol: 48.94% versus 51.1%, P = 0.89). The multiple pregnancy rate and the live birth rate were also similar between the groups. No significant differences in gestational age, birthweight or proportion of male/female newborns were observed. The concentration of cfDNA in the blastocyst culture medium was not associated with IVF outcome.
CONCLUSIONS: Large-scale randomized controlled trials are required to confirm these preliminary results.

UNASSIGNED: This work investigates ICSI outcome between LASER Artificial Shrinkage (LAS) and Micro-Needle Artificial Shrinkage (MNAS) before vitrification.
UNASSIGNED: Four hundred and nine women were included in the study; which were randomly divided into two groups according to the technique used for artificial shrinkage step of the blastocyst before vitrification: in the first group, Laser beam was used while in the second group the micro-needle was used. Ovarian stimulation was done before the ICSI procedure either by long, short or antagonist protocol.
UNASSIGNED: The statistical analysis of our study revealed that there was no statistically significant difference between the two groups regarding age, number of cases, AMH, Basal FSH, BMI, male factor, usage ovarian stimulation protocol, high quality blastocysts, the mean number of transferred embryos. While, there was a statistically significant difference between two groups after thawing with p-value < .001 in favor of the LAS method regarding the morphology of originally high quality blastocysts, blastocysts healthiness (not degenerated), pregnancy rate, the implantation rates.
UNASSIGNED: LASER artificial shrinkage of human blastocysts is a promising technology that could be implemented on a wider basis to improve ART practice, as our study revealed that the usage of LASER pulse for artificial shrinkage of blastocysts before vitrification has better implantation rate as well as better chemical and clinical pregnancy rate in comparison to the usage of micro-needle artificial shrinkage of blastocysts before vitrification. There is a statistically significant difference regarding live birth rate being more in the LASER group as compared to needle group, also the number of twins ether identical or non-identical are larger in laser group than in needle group but with no statistically significant difference. Clinical trials.gov ID: NCT04125017.

To present an effective approach to trophectoderm biopsy for blastocysts of different stages and characteristics by mechanical blunt dissection (MBD).
Stepwise demonstration with still pictures and operational video clips to explain tips and tricks for trophectoderm biopsy. (This demonstration was approved by the Reproductive Study Ethics Committee at Shengjing Hospital of China Medical University.) SETTING: In vitro fertilization laboratory.
Patients who underwent preimplantation genetic testing.
The illustrated techniques of blastocyst trophectoderm biopsy using micromanipulation methods include artificial shrinkage, zona pellucida drilling, injecting media from the drilling, aspiration of trophectoderm cells into the biopsy pipette (outer diameter 27 μm for fully expanded blastocysts and peanut-shaped hatching blastocysts; outer diameter 20 μm for 8-shaped hatching and hatched blastocysts), detachment of the trophectoderm cells by laser pulse combined with MBD (performed using the rims of the biopsy and holding pipettes), and release of the biopsy fragment.
Successful biopsy rate and survival after warming.
Our biopsy strategy does not involve assisted hatching on day-3 or day-4 embryos, which can leave the embryo undisturbed in culture up to the expanded blastocyst stage. Notably, this approach demonstrates several noteworthy advantages for sampling blastocysts of different stages and characteristics, and it maintains a desirable successful biopsy rate (95.4%, n = 1,872) and survival rate after warming (100%, n = 440). The MBD method may reduce thermal damage because fewer laser pulses are used, compared with the traditional laser-only biopsy techniques. For noncollapsed blastocysts after artificial shrinkage, the strategy of injecting medium from the zona pellucida drilling helps to separate the trophectoderm cells from the zona pellucida, thus facilitating the biopsy procedure. For peanut-shaped hatching blastocysts, this approach could provide better control over the aspiration of trophectoderm cells into the biopsy pipette. Especially if the inner cell mass is herniating from the zona pellucida, the trophectoderm biopsy can be performed away from the inner cell mass to avoid damaging it. In addition, the MBD approach combined with the biopsy pipette (outer diameter 20 μm) can effectively control the target number of trophectoderm cells, thus simplifying the process of obtaining a biopsy from a hatched blastocyst.
Our biopsy approach demonstrates several noteworthy advantages. Considering its benefits and the simplicity of its execution, this systematic biopsy method for blastocysts of different stages and characteristic can be widely applied.

Embryo vitrification is increasingly used in IVF. Artificial shrinkage (collapse) before vitrification has been proposed to maximise blastocyst survival after warming. However, its effectiveness on blastocyst survival rate and vitrified-warmed blastocyst transfer cycle outcome remains to be confirmed. Therefore, we performed a systematic MEDLINE search according to PRISMA guidelines on all articles published up to April 2018 and related to human blastocyst collapse before vitrification using the following keywords: (i) blastocyst; (ii) collapse; (iii) artificial shrinkage; and (iv) vitrification. The following outcomes were analysed and included in the meta-analysis: (i) blastocyst survival rate after warming; (ii) implantation rate; (iii) clinical pregnancy rate; and (iv) live birth rate after vitrified-warmed blastocyst transfer (commonly named frozen-thawed blastocyst transfer). Eight articles were included. Briefly, blastocyst survival (OR 5.04, 95% CI 2.43-10.46) and clinical pregnancy rate (OR 1.87, 95% CI 1.26-2.77) were significantly higher in collapse than in control group. However, implantation rate (OR 2.50, 95% CI 0.67-9.28) and live birth rate (OR 1.35, 95% CI 0.88-2.09) were comparable in both groups. In conclusion, this systematic review and meta-analysis suggests that artificial shrinkage before blastocyst vitrification improves survival and clinical pregnancy rate, but not implantation or live birth rate. Further randomised studies are warranted to improve the level of evidence and confirm these findings.

OBJECTIVE: Microsuction (MS) is a technique for mechanically emptying fluid from blastocele using a microneedle. In this study, we evaluated the improvement in clinical and neonatal outcomes of vitrified blastocyst transfer programs when MS of blastocelic fluid was used before vitrification.
METHODS: This was a retrospective study based on data collected between March 2014 and August 2016. A total of 317 blastocysts obtained from 211 patients were analyzed. The blastocelic fluid of expanded blastocysts was aspirated completely, and blastocysts were collapsed prior to vitrification. Clinical and neonatal outcomes of warmed blastocysts were compared.
RESULTS: The survival rate of the MS blastocyst was significantly higher compared with the nontreatment control (98.7% vs 89.3%, OR: 9.34, 95% CI: 2.35-36.8, P < 0.01). The rates of implantation and live birth were higher in the MS group than in the control group, but the differences were not significant. There were no differences in gestational age, birthweight, proportion of male babies, rates of cesarean section, and congenital abnormalities.
CONCLUSIONS: The MS procedure improved blastocyst survival and had little effect on further embryo development after warming.

BACKGROUND: In recent years, single blastocyst transfer combined with vitrification has been applied widely, which can maximize the cumulative pregnancy rate in per oocyte retrieval cycles and minimize the multiple pregnancy rate. Thus, the guarantee for these is the effectiveness of vitrified blastocyst. Studies has shown that AS of the blastocoel cavity prior to vitrification can reduce injuries, increase the thawed blastocyst survival rate and implantation rate. Several AS methods have been established. However, only a few studies have compared the effectiveness and safety of these AS methods. In this study, we aimed to compare the clinical outcomes and neonatal outcomes in FET cycles with single blastocyst that were artificially shrunk before vitrification by either LAS or MNAS method.
METHODS: A retrospective comparative study of FET cycles in infertile patients which were at our clinic between January 2013 and December 2014. These FET cycles were divided into two groups by the shrinking methods used before vitrification and the clinical and neonatal outcomes were assessed.
RESULTS: There were no statistically differences in blastocyst survival rates (95.40% vs 94.05%, P > 0.05) between the LAS and MNAS groups. However, compared with MNAS, LAS improved the warmed blastocyst implantation/clinical pregnancy rate (60.82% vs 54.37%, P < 0.05), live birth rate (50.43% vs 45.22%, P < 0.05) and also increased the monozygotic twin rate (4.07% vs 1.73%, P < 0.05). There were no differences in the average gestational weeks (38.83 ± 1.57 vs 38.74 ± 1.75), premature birth rate (0.30% vs 0.49%), average birth weight (3217.89 ± 489.98 g vs 3150.88 ± 524.03 g), low birth weight rate (5.60% vs 8.63%) and malformation rate (0.59% vs 0.48%) (P > 0.05).
CONCLUSIONS: No significant differences in neonatal outcomes were observed, while in clinical outcomes, LAS improved the warmed blastocyst implantation/clinical pregnancy rate and live birth rate markedly, there was also an increased risk of monozygotic twin pregnancies.

OBJECTIVE: Blastocysts contain a large amount of fluid in the blastocoel, which may pose a risk for ice crystal formation during vitrification. This study aimed to evaluate the effectiveness of laser-induced artificial shrinkage of blastocoel before vitrification on clinical outcome.
METHODS: Patients were divided into two groups: a control group with untreated, expanded blastocysts (n = 115) and a study group with blastocoel artificially eliminated by a laser pulse prior to vitrification (n = 309). Blastocyst survival, clinical pregnancy, and implantation rates were compared.
RESULTS: The survival rate was significantly higher in the study group compared with the control group (97.3 and 74.9 %, respectively; p > 0.01). The clinical pregnancy and implantation rates of the study group were significantly higher (p < 0.01) than that of the control group (clinical pregnancy, 67.2 vs. 41.1 %; implantation, 39.1 vs. 24.5 %.
CONCLUSIONS: This study demonstrated that the removal of blastocoel fluid before vitrification by laser pulse of in vitro-produced human blastocysts significantly improves blastocyst survival, clinical pregnancy, and implantation rates.

OBJECTIVE: This study aims to compare implantation, pregnancy, and delivery rates in frozen transfer cycles with blastocysts that were vitrified either with artificial shrinking (AS group) or without (NAS group).
METHODS: Retrospective comparative study of artificial shrinking of blastocysts prior to vitrification and frozen embryo transfer cycles in infertile patients undergoing frozen embryo transfer (FET) was done at the Humanitas Fertility Center between October 2009 and December 2013. Main outcome measure(s) were implantation (IR), pregnancy (PR), and delivery rates (DR) between the two groups.
RESULTS: A total of 1028 consecutive warming blastocyst transfer cycles were considered. In 580 cycles (total of 822 blastocysts), artificial shrinking was performed prior to vitrification (AS group), while in the remaining 448 cycles (total of 625 blastocysts), the artificial shrinking was not performed (NAS group). There were no differences in patient age (36.4 ± 3.7 vs. 36.3 ± 3.9) and number of embryos transferred (1.41 ± 0.49 vs. 1.38 ± 0.50) between groups. The IR, PR, and DR in the AS group were significantly higher (p < 0.05) than in the NAS group (29.9 vs. 23.0 %, 36.3 vs. 27.9 %, and 26.5 vs. 18.1 %, respectively).
CONCLUSIONS: Performing AS of blastocysts prior to vitrification appears to improve implantation, pregnancy, and delivery rates probably related to a decreased risk of ultrastructural cryodamages, plausible when cryopreserving expanded blastocysts.

OBJECTIVE: The goal of this study was to ascertain optimal assisted hatching (AH) method in frozen embryo transfer. We compared the effect of depending on whether mechanical or laser-AH was performed before or after the vitrification of embryo development rate and blastocyst cell numbers.
METHODS: In order to induce superovulation, pregnant mare\'s serum gonadotropin followed by human chorionic gonadotropin were injected into 4- to 5-week-old female mice. 2-cell embryos were then collected by flushing out the oviducts. The Expanded blastocysts were recovered after the collected embryos were incubated for 48 hours, and were then subjected to artificial shrinkage (AS) and cross-mechanical AH (cMAH) or quarter-laser zona thinning-AH (qLZT-AH) were carried out using the expanded blastocysts before or after vitrification. After 48 hours of incubation, followed by vitrification and thawing (V-T), and blastocysts were fluorescence stained and observed.
RESULTS: The rate of formation of hatched blastocysts after 24 and 72 hours of incubation was significantly higher in the AS/qLZT-AH/V-T group than in the other groups (p<0.05). The cell number of the inner cell mass was higher in AS/V-T/non-AH and AS/V-T/cMAH groups than those of others (p<0.05). In the control group, the number of trophectoderm and the total cell number were higher than in the AS-AH group (p<0.05).
CONCLUSIONS: The above results suggest that AS and AH in vitrification of expanded blastocysts lead to the more efficient formation of hatched blastocysts in mice.

OBJECTIVE: What is the effect of artificial shrinkage by laser-induced collapse before vitrification on the implantation potential after transfer of vitrified-warmed blastocysts?
CONCLUSIONS: The artificial shrinkage by laser-induced collapse did not significantly increase the implantation rate per transferred collapsed blastocyst (37.6%) compared with non-collapsed blastocysts (28.9%) [odds ratio (OR): 1.48, 95% confidence interval (CI): 0.78-2.83].
BACKGROUND: Retrospective studies have demonstrated that artificial shrinkage of the blastocyst prior to vitrification can have a positive effect on blastocyst survival after warming. A recent study found a similar survival rate but higher implantation rate for collapsed blastocysts. So far, no randomized controlled trial has been conducted to investigate the implantation potential of collapsed blastocysts.
METHODS: Prospective randomized trial. Patients were recruited from December 2011 until April 2014 and warming cycles were included until July 2014. Patients were randomized in the fresh cycle if blastocysts were available for vitrification and were allocated to the study or control arm according to a computer-generated list. In the study group, blastocysts underwent laser-induced collapse before vitrification. In the control group, blastocysts were vitrified without collapsing.
METHODS: In total, 443 patients signed informed consent and 270 patients had blastocysts vitrified. One-hundred and thirty-five patients were allocated to the study group and 135 to the control group. Sixty-nine patients from the study group and 69 from the control group returned for at least one warming cycle in which 85 and 93 blastocysts were warmed in the first cycle, respectively. Primary outcome was implantation rate per embryo transferred in the first warming cycle. Secondary outcomes were survival and transfer rates, blastocyst quality after warming, clinical pregnancy rate and implantation rate per warmed blastocyst. Blastocysts were vitrified-warmed one by one using closed vitrification and one or two blastocysts were transferred per warming cycle.
RESULTS: We calculated that the group sample sizes of 80 embryos in the collapse group and 80 embryos in the control group were needed to achieve 80% power to detect a difference between the group proportions of +20% with P < 0.05. In the study group, 69 first warming cycles resulted in 69 transfers with 1.2 blastocysts (n = 85) transferred. In the control group, an average of 1.3 blastocysts (n = 83) were transferred in 67 out of 69 warming cycles. Implantation rates per embryo transferred in the first warming cycle were not different between both groups (38 versus 29%, OR: 1.48; 95% CI: 0.78-2.83), neither was the implantation rate per warmed embryo (38 versus 26%, OR: 1.74; 95% CI: 0.92-3.29). When all warming cycles were considered (n = 135 in each group), survival rate after collapse was significantly higher compared with the control group (98.0 versus 92.0%, OR: 4.25; 95% CI: 1.19-15.21). Furthermore, a higher percentage of high-quality blastocysts (36.3 versus 23.5%, OR: 1.86; 95% CI: 1.12-3.08) and hatching blastocysts (19.2 versus 5.4%, OR: 4.18; 95% CI: 1.84-9.52) were found compared with the control group.
CONCLUSIONS: The study lasted more than 2.5 years since fewer patients than expected returned for a warming cycle because of the high ongoing pregnancy rates in the fresh IVF/ICSI cycle.
CONCLUSIONS: Although no significant higher implantation rate was found after collapse, the better survival and post-warm embryo quality convinced us to recognize a clinical benefit of artificial shrinkage and to implement it in routine vitrification practice.
BACKGROUND: NCT01980225, www.clinicaltrials.gov. The first patient was included November 2011 and the study was registered October 2013.