Technical note: Mice produced by intracytoplasmic sperm injection using a modified conventional method1

Liu, J.; Lee, G. Y.; Lawitts, Joel; Toner, Mehmet; Biggers, John D.

doi:10.2527/jas.2012-5136

Cited by 7 publications

(4 citation statements)

References 10 publications

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“…ICSI was applied using a modified conventional method [8]. Twenty micro liters of sterile distilled water were added to each dried sample (a thin film) to rehydrate to its original volume.…”

Section: Methodsmentioning

confidence: 99%

Live Pups from Evaporatively Dried Mouse Sperm Stored at Ambient Temperature for up to 2 Years

et al. 2014

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The purpose of this study is to develop a mouse sperm preservation method based on evaporative drying. Mouse sperm were evaporatively dried and stored at 4°C and ambient temperature for 3 months to 2 years. Upon rehydration, a single sperm was injected into a mature oocyte to develop into a blastocyst after culture or a live birth after embryo transfer to a recipient female. For the samples stored at 4°C for 3, 6, 12, 18, and 24 months, the blastocyst formation rate was 61.5%, 49.1%, 31.5%, 32.2%, and 41.4%, respectively. The blastocyst rate for those stored at ambient temperature (∼22°C) for 3, 6, 12, and 18 months was 57.8%, 36.2%, 33.6%, and 34.4%, respectively. Fifteen, eight and three live pups were produced from sperm stored at room temperature for 12, 18, and 24 months, respectively. This is the first report of live offspring produced from dried mouse sperm stored at ambient temperature for up to 2 years. Based on these results, we suggest that evaporative drying is a potentially useful method for the routine preservation of mouse sperm.

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Section: Methodsmentioning

confidence: 99%

Live Pups from Evaporatively Dried Mouse Sperm Stored at Ambient Temperature for up to 2 Years

et al. 2014

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“…ICSI was performed as previously described. 42 Briefly, sperm heads and tails were separated by sonication. A single sperm head was picked up by an injection pipette and inserted into an oocyte.…”

Section: Methodsmentioning

confidence: 99%

On-chip oocyte denudation from cumulus–oocyte complexes for assisted reproductive therapy

et al. 2018

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Human infertility can be treated using assisted reproductive technology (ART) such as intracytoplasmic sperm injection (ICSI). But current ART techniques suffer from multiple cumbersome processes requiring technically skilled personnel. Microfluidics technologies offer unique opportunities to streamline ART procedures, reduce stress imposed upon gametes and embryos, and minimize the operator-to-operator variability. However, there have been no automated and continuous processing systems that can reduce the dependence on well-trained embryologists to obtain ICSI-ready oocytes from patients. In this study, using mouse models, we developed a microfluidic device to denude oocytes from the surrounding cumulus-corona cell mass, facilitating the evaluation of oocyte quality and the injection of sperm. Enzyme-treated cumulus-oocyte complexes pass through a series of jagged-surface constriction microchannels of optimized geometries. The jagged inner wall of constriction channels facilitates to strip off the cumulus-corona cell mass. Oocytes that were denuded by the device showed comparable fertilization and developmental competence compared with mechanical pipetting. The device developed in this study achieves the automation of manual process for oocyte denudation in a continuous flow, as well as improving standardization and ease-of-use. Our denudation-on-a-chip approach requires inexpensive and simple equipment, which represents one step forward towards improving the accessibility and affordability of assisted reproductive therapy.

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“…To realize ICSI, many techniques have been developed such as conventional ICSI [2,9,10], piezo-assisted ICSI [4,11,12], and the Ros-drill technique [13]. Conventional ICSI is commonly implemented in humans and domestic animals where the rupture of the oolemma is achieved by largely deforming the oocyte.…”

Section: Introductionmentioning

confidence: 99%

“…However, mercury causes concerns of toxicity and contamination to the manipulated oocytes [15][16][17]. Although FC-77 can be used as a substitute for mercury, its damping effect is not as satisfactory as mercury [9]. To avoid using mercury, a piezo-driven technique was reported for the puncture of zebrafish embryos [18].…”

Section: Introductionmentioning

confidence: 99%

The Development of Piezo-Driven Tools for Cellular Piercing

Pan

Chen

2016

Applied Sciences

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Conventionally, intracytoplasmic sperm injection (ICSI) in the mouse is conducted with piezo-drills that use a droplet of mercury for damping. The use of mercury causes concerns of toxicity and contamination. Although Fluorinert can be used as a substitute for mercury to reduce piezo-drill's lateral vibration, the damping effect is not as satisfactory as mercury. In this work, a modified piezo-drill without using mercury was developed for the cellular piercing of mouse oocytes. Experimentally, appropriate parameters of driving voltage and frequency were obtained for the penetration of the zona pellucida of mouse oocytes. Furthermore, the lateral vibration of the injection pipette is lower than 1 µm in deionized water, which is not observable at 400 magnificence. With the piezo-drill without using mercury, the system performs the cellular piercing of mouse oocytes with a maximum cleavage rate of 94.7% (n = 117).

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Technical note: Mice produced by intracytoplasmic sperm injection using a modified conventional method1

Cited by 7 publications

References 10 publications

Live Pups from Evaporatively Dried Mouse Sperm Stored at Ambient Temperature for up to 2 Years

Live Pups from Evaporatively Dried Mouse Sperm Stored at Ambient Temperature for up to 2 Years

On-chip oocyte denudation from cumulus–oocyte complexes for assisted reproductive therapy

The Development of Piezo-Driven Tools for Cellular Piercing

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