Vitrification of immature bovine oocytes in protein-free media: The impact of the cryoprotectant treatment protocol, base medium, and ovary storage

Somfai, T.; Hirao, Yuji

doi:10.1016/j.theriogenology.2021.05.029

Cited by 7 publications

(3 citation statements)

References 42 publications

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“…Nevertheless, after vitrification at the GV stage, surviving oocytes showed reduced competence for cleavage and subsequent embryo development unlike zygotes vitrified by the same protocol. These results are in agreement with our previous reports in pigs [8,11,21,27] and cattle [22,28]. Previously we demonstrated that our vitrification protocol does not impair the ability of GV-stage oocytes to undergo maturation and subsequent fertilization [11][12][13]26].…”

Section: Plos Onesupporting

confidence: 94%

Vitrification of porcine immature oocytes and zygotes results in different levels of DNA damage which reflects developmental competence to the blastocyst stage

et al. 2023

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The present study investigated the effects of vitrification of porcine oocytes either at the immature Germinal Vesicle (GV) stage before in vitro maturation (GV-stage oocytes) or at the pronuclear stage after in vitro maturation and fertilization (zygotes) on DNA integrity in relevance with their subsequent embryo development. Vitrification at the GV stage but not at the pronuclear stage significantly increased the abundance of double-strand breaks (DSBs) in the DNA measured by the relative fluorescence after γH2AX immunostaining. Treatment of GV-stage oocytes with cryoprotectant agents alone had no effect on DSB levels. When oocytes were vitrified at the GV stage and subjected to in vitro maturation and fertilization (Day 0) and embryo culture, significantly increased DSB levels were detected in subsequent cleavage-stage embryos which were associated with low cell numbers on Day 2, the upregulation of the RAD51 gene at the 4–8 cell stage (measured by RT-qPCR) and reduced developmental ability to the blastocyst stage when compared with the non-vitrified control. However, total cell numbers and percentages of apoptotic cells (measured by TUNEL) in resultant blastocysts were not different from those of the non-vitrified control. On the other hand, vitrification of zygotes had no effect on DSB levels and the expression of DNA-repair genes in resultant embryos, and their development did not differ from that of the non-vitrified control. These results indicate that during vitrification GV-stage oocytes are more susceptible to DNA damages than zygotes, which affects their subsequent development to the blastocyst stage.

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Section: Plos Onesupporting

confidence: 94%

Vitrification of porcine immature oocytes and zygotes results in different levels of DNA damage which reflects developmental competence to the blastocyst stage

et al. 2023

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“…In Cryotop vitrification of bovine immature oocytes, the use of EG/PG‐based VS instead of EG/DMSO‐based VS resulted in a higher IVM outcome (90% vs. 76%), but a comparable blastocyst yield (10% vs. 14% per matured oocytes; Faheem et al, 2015 ). Recently, Somfai and Hirao ( 2021 ) reported that the replacement of DMSO with PG in protein‐free VS with a few additional modifications resulted in a statistically comparable blastocyst yield from post‐warmed bovine immature oocytes (12% vs. 8%). In the above experiment, the positive effect of the EG/PG‐based protein‐free VS was clearly demonstrated in the total cell number of resulting blastocysts (118.1 vs. 56.5 cells).…”

Section: Application To Immature Oocytesmentioning

confidence: 99%

Cryodevices developed for minimum volume cooling vitrification of bovine oocytes

Hochi

2022

Animal Science Journal

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Unfertilized bovine oocytes can be efficiently cryopreserved only when an extremely rapid cooling rate (>20,000°C/min) is applied to oocytes with a very limited amount of surrounding vitrification solution. This protocol is defined as minimum volume cooling (MVC) vitrification. Various types of cryodevices, such as open pulled straw, Cryoloop, and Cryotop, have been developed to accelerate the cooling efficacy. Furthermore, hollow fibers with nano‐scale pores, triangle nylon mesh sheets, and multilayer silk fibroin sheets have been optimized for the loading of large quantities of oocytes and/or the subsequent removal of excess vitrification solution, without requiring skillful operation to transfer individual oocytes using fine capillaries. This article provides an up‐to‐date review of cryodevices suitable for the MVC vitrification of bovine oocytes at the immature (germinal vesicle‐) and mature (metaphase II‐) stages.

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“…Among the method of ultra-rapid freezing, this method is less costly, more straightforward, and most used. Plunging involves placing a biological sample either with or without a container directly in liquid cryogens [71,[213][214][215][216][217][218][219][220][221][222][223][224][225][226][227]; however, compared with the conventional rapid freezing method, there are four effective conditions to improve the cooling rate: High entry velocity of the sample into a cryogen (minimum 1 m/s) [228][229][230][231]; High surface-to-volume ratio of the sample (e.g., a copper mesh or a device such as a cryoloop or cryotop) [120][121][122]125,216,217,222,232,233]; The lowest possible temperature of cryogen (e.g., nitrogen slush or liquid helium) [232,[234][235][236][237][238][239]; Stirring the cryogen in a container to decrease temperature gradients [235,240].…”

Section: Plunge Freezingmentioning

confidence: 99%

Technologies for Vitrification Based Cryopreservation

Amini

Benson

2023

Bioengineering

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Cryopreservation is a unique and practical method to facilitate extended access to biological materials. Because of this, cryopreservation of cells, tissues, and organs is essential to modern medical science, including cancer cell therapy, tissue engineering, transplantation, reproductive technologies, and bio-banking. Among diverse cryopreservation methods, significant focus has been placed on vitrification due to low cost and reduced protocol time. However, several factors, including the intracellular ice formation that is suppressed in the conventional cryopreservation method, restrict the achievement of this method. To enhance the viability and functionality of biological samples after storage, a large number of cryoprotocols and cryodevices have been developed and studied. Recently, new technologies have been investigated by considering the physical and thermodynamic aspects of cryopreservation in heat and mass transfer. In this review, we first present an overview of the physiochemical aspects of freezing in cryopreservation. Secondly, we present and catalog classical and novel approaches that seek to capitalize on these physicochemical effects. We conclude with the perspective that interdisciplinary studies provide pieces of the cryopreservation puzzle to achieve sustainability in the biospecimen supply chain.

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Vitrification of immature bovine oocytes in protein-free media: The impact of the cryoprotectant treatment protocol, base medium, and ovary storage

Cited by 7 publications

References 42 publications

Vitrification of porcine immature oocytes and zygotes results in different levels of DNA damage which reflects developmental competence to the blastocyst stage

Vitrification of porcine immature oocytes and zygotes results in different levels of DNA damage which reflects developmental competence to the blastocyst stage

Cryodevices developed for minimum volume cooling vitrification of bovine oocytes

Technologies for Vitrification Based Cryopreservation

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