الفهرس 
REVIEW OF LITERATUREOnly 14 pages are availabe for public view
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Abstract
SUMMARY AND CONCLUSION
This study was carried out to investigate the effect of BCB, culture media, gonadotropins, proteins, growth factors and hyaluronic acid on in-vitro maturation rate of buffalo oocytes, effect of fertilization media on fertilization rate and effect of culture media on embryo development. The study was conducted through 2017 -2019 in the Department of Animal Reproduction and Artificial Insemination, Veterinary Research Division. National Research Centre, Dokki, Egypt. Buffalo Ovaries were collected from local abattoir (Shubra El-Khaimah, Qualyubia, Egypt).
The results obtained from this study could be summarized as follows:
Immature oocytes quality
Aspiration method led to a good recovery rate of oocytes and a good proportion of oocytes suitable for maturation. Whereas the mean percent of grade A, B, C and D were 55.05 ± 0.54, 22.46 ± 0.39, 13.29 ± 0.61 and 9.4 ± 0.51, respectively.
Impact of immature oocyte segregation by BCB into BCB+ and BCB- on cumulus cell expansion and nuclear maturation
oocytes were colored with blue (BCB+) was significantly higher than oocytes that refused to accept the dye (BCB-).
BCB+ and control group achieved significantly (P<0.05) higher percentage of G3 (full cumulus-cell expansion), and percentage of 1st polar body (Cytoplasmic and nuclear maturation) compared with BCB-.
First Part: In-vitro maturation
First experiment (Effect of Maturation Media on In-Vitro Maturation Rate of Buffalo Oocytes)
This study was carried out to investigate the effect of TCM-199, CR1aa and DMEM media on maturation rate of buffalo oocytes.
The main results of the experiment: Indicated that buffalo oocytes incubated in TCM-199 show significantly higher percentage of both cytoplasmic and nuclear maturation (P< 0.05) followed by that incubated in CR1aa and DMEM media, respectively.
Second experiment (Effect of Supplementing TCM-199 with Hormones on Maturation Rate of Buffalo Oocytes)
This study was carried out to investigate the effect of using FSH and/or eCG on maturation rate of buffalo oocytes.
The main results of the experiment include Supplementation of maturation medium with eCG with or without FSH achieved significantly (P<0.05) an increase of full cumulus-cell expansion (G3) percentage and percentage of 1st polar body (Cytoplasmic and nuclear maturation). Regarding percentage of moderate cumulus-cell expansion (G2) IVM medium supported with combination of FSH and eCG obtained the highest percentage (P<0.05) compared with IVM medium supported with FSH or eCG. On the other hand, there is no significant difference between slight cumulus cell expansions (G1) percentage of buffalo oocytes cultured in IVM media provided with FSH with or without eCG. Whereas addition of FSH to IVM medium led to significantly (P<0.05) higher percentage of G0 compared with adding eCG with FSH.
Third experiment (Effect of adding protein or PVA to maturation medium on maturation rate of buffalo oocytes)
This experiment was carried out to investigate the effect of using FCS, BSA or PVA on maturation rate of buffalo oocytes.
The main results of the experiment indicated: Adding FCS as a supplement to IVM medium improved (P< 0.05) the percentage of cytoplasmic and nuclear maturation of buffalo oocyte when compared with using BSA or PVA. While there is no significant difference between moderate cumulus cell (G2) expansion percentage of buffalo oocytes cultured in IVM medium supplemented with FCS, BSA and PVA. Whereas, adding FCS or BSA to IVM medium significantly decreased percentage of slight cumulus-cell expansion (G1) compared with adding PVA to IVM.
Fourth experiment (Effect of adding growth factors to maturation medium on maturation rate of buffalo oocytes)
This experiment was carried out to investigate the effect of using IGF-1 or EGF on maturation rate of buffalo oocytes.
The main results of the experiment indicated: Enrichment IVM (TCM-199) medium with EGF increased (P<0.05) percentage of G3 and 1st polar body extruded when compared with IGF-1 or without addition. Whereas there is no significant difference between moderate cumulus cell expansion (G2) percentage of buffalo oocytes cultured in IVM medium supplemented with/without growth factor. Regarding percentage of slight cumulus-cell expansion (G1) IVM medium without addition growth factors (control) obtained high (P<0.05) percentage compared with IVM medium supported with EGF, there is no significant difference when adding IGF-1 to IVM medium than control maturation medium or maturation medium with EGF. Control maturation medium or maturation medium supplemented with IGF-1 increased (P<0.05) percentage of G0 compared to EGF added to maturation medium.
Fifth experiment (Effect of adding different concentrations of hyaluronic acid to maturation medium on maturation rate of buffalo oocytes)
This experiment was carried out to investigate the effect of using 0, 2.5, 5.0 or 10 µg/mL hyaluronic acid on maturation rate of buffalo oocytes.
The main results of the experiment indicated: The percentage of full cumulus-cells expansion and % nuclear maturation was significantly increased by adding 2.5µg/mL HA to maturation medium compared with other groups, there is no significant difference between control group and group treated with 5µg/mL in nuclear maturation, while control group was better in percentage of full cumulus-cells expansion. Adding 10 µg/mL HA to maturation medium inhibit significantly cytoplasmic maturation (P< 0.05). So, it increases the percentage of non-mature oocytes and increased (P< 0.05) percentage of G0 cumulus cell expansion compared with other groups.
Part 2: Effect of Fertilization Media on In-Vitro Fertilization Rate of Matured Buffalo Oocytes.
This study was carried out to investigate the effect of CR1aa, TALP, and BO fertilization media on fertilization rate of matured buffalo oocytes.
The main results of the experiment indicted that: CR1aa fertilization medium showed significant superiority over both the TALP and BO media. TALP fertilization medium showed significant improvement in fertilization rate compared with BO medium
Part 3: Effect of Culture Media on In-Vitro Culture Rate of Buffalo Zygotes
This study was carried out to investigate the effect of SOF and CR1aa culture media on cleavage and embryo development rate of buffalo zygotes.
The main results of the experiment indicated: that there were no significant differences between SOF and CR1aa culture media on cleavage, Morula, and Blastocysts rate of buffalo zygotes.
But there is a non-statistical superiority of the ٍSOF media over the CR1aa media in percentage of cleavage. Morula, and Blastocysts of buffalo zygotes.