Selection of patients for natural cycle in vitro fertilization combined with in vitro maturation of immature oocytes

The main objectives of this study is the separation of X from Y bearing epididymal spermatozoa of local buck by swim-up, and the use of this spermatozoa for in vitro fertilization to determine the percentage of produced male and female embryos. The sex of produced embryo was identified by polymerase chain reaction. Testis of the local buck were obtained from Al-Shu'alah abattoir and the epididymal spermatozoa were harvested from the cauda by and submitted to in vitro maturation prior to separation of X from Y bearing spermatozoa and prior to their use for in vitro fertilization. For the separation of epididymal spermatozoa, swim-up technique was used with centrifugation at 200×g or 300×g. The centrifugation at 200×g showed that 41.84±1.39 % of spermatozoa were detected in the supernatant while the precipitate contained 50.69±0.71 and the mean of the sperm lost was 7.65±0.93. After centrifugation, spermatozoa in the supernatant were used for in vitro fertilization of matured oocytes. The sex of in vitro produced goat embryos was determined by polymerase chain reaction using specific primers to detect of SRY gene. The percentage of total goat embryos obtained after in vitro fertilization by sperms selected using swim-up at centrifugation force of 200×g recorded 79.66 % male embryos while female embryos recorded only 20.33 %. At the end, the results showed the ability of selection male embryos in caprine by application of swim-up technique on epididymal spermatozoa with centrifugation at 200×g.

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CRYOPRESERVATION OF SHEEP OOCYTES AND EMBRYOS USING LOCAL, MODIFIEID VITRIFICATION TOOL

IRAQI JOURNAL OF AGRICULTURAL SCIENCES ◽

10.36103/ijas.v50ispecial.192 ◽

2019 ◽

Vol 50 (Special) ◽

Author(s):

Atiyah & et al.

Keyword(s):

In Vitro Fertilization ◽

Embryo Development ◽

In Vitro Maturation ◽

Cost Effective ◽

Normal Morphology ◽

Vitro Fertilization ◽

Immature Oocytes ◽

Cell Embryo ◽

First Time

The present study was aimed to cryopreserve mature, immature oocytes and in vitro produced embryos in Iraqi sheep using vitrification technique by local, simple and cost effective vitrification tool. This tool is an innovative straw called vitripeace invented, designed and used for the first time. Immature oocytes were aspirated from ovaries of slaughtered ewes and subjected to in vitro maturation and in vitro fertilization programs.The immature, mature oocytes and embryos were vitrified using Vitripeace tools, then thawed and assessed for the morphology and viability. The results revealed non-significant effect of time on viability (%) and normal morphology (%) of vitrified immature and mature oocytes for post-thawing and 2 hours post-thawing. The results showed significant (P<0.05) reduction in the viability (%) of 2 cell embryo namely 88.89% and 77.78 % for post-thawing and two hours post-thawing respectively. The results revealed a significant (P<0.05) reduction on normal morphology of 1 cell embryo namely 88.24 % and 76.47 % for post-thawing and two hours post-thawing respectively. Significant (P<0.05) differences in the percentage of normal morphology were found at post-thawing period for all stages of embryo development which were 90.74%, 88.31, 88.24 and 83.33 for immature , mature oocytes, 1 cell and 2 cell embryos, respectively while no significant differences in the viability at post-thawing period among all stages of embryo development. It was concluded that, successful vitrification of oocytes and embryos was resulted using Vitripeace which was novel, simple and cost effective vitrification tool.

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