Influence of serum source on buffalo in vitro embryonic development with reference to heat inactivation

Mahmoud, Karima Gh. M.; Nawito, M.

doi:10.1016/j.livprodsci.2005.02.005

Cited by 2 publications

(1 citation statement)

References 26 publications

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“…Further, similar result to earlier report has been demonstrated elsewhere (Schellander et al, 1990). There are different sources for supplemented sera such as FBS (Kobayeshi et al, 1994;Nandi, 1998;Mahmoud and Nawito, 2005;Das et al, 2006;Talukder et al, 2008), OCS (Schellander et al, 1990;Singha et al, 2015), steer serum (Roy et al, 1968;Nandi et al, 2001) and superovulated cow serum (Boediono et al, 1994). Although, there are reports to use different sera sources for supplementation in maturation medium, comparison was made within FBS, OCS and BSA only in the present study.…”

Section: Experiments 3 Determination Of An Effective Protein Supplemementioning

confidence: 87%

Determination of an Effective Media and Its Hormone and Protein Supplemenntation for in Vitro Maturation of Oocytes of Indigenous Zebu Cows

Rahman¹,

Morshed²,

Juyeana³

et al. 2018

Bangl. J. Vet. Med.

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In vitro maturation (IVM) of oocytes is the first important step for successful in vitro embryo production of any mammalian species. The objectives of the present study were to determine an effective basic medium and its hormone and protein supplementation for IVM of oocytes of indigenous zebu cows. The oocytes were derived from ovaries of locally slaughtered cows after aspiration of follicle. The oocytes were cultured in medium for 24 hrs at 38.5ºC with 5% CO2 in humidified air for maturation. The maturation of oocytes was evaluated by examining the presence of first polar body extrusion in denuded oocytes under inverted microscope. To determine an effective basic medium, the oocytes were cultured in fetal bovine serum (FBS) supplemented tissue culture medium (TCM), modified synthetic oviduct fluid (mSOF) and Tyrodes albumin lactate pyruvate (TALP) medium. The maturation rate was significantly higher (74±4.2) in TCM medium than that of TALP medium (58.2±6.2). To determine an effective hormone supplementation for maturation medium, the oocytes were cultured in either in follicle stimulating hormone (FSH) or gonadotrophin supplemented TCM. The maturation rate of oocytes was significantly (p>0.05) higher (73.3±4.0) in FSH supplemented medium than that of gonadotrophin supplemented counterpart (60.2±6.6). To determine an effective protein supplementation, the oocytes were cultured in FBS, oestrus cow serum (OCS) and bovine serum albumin (BSA) supplemented TCM 199. The maturation rate of oocytes were 73.0±5.9, 71.1±2.8, and 62.5±9.4 in medium supplemented with FBS, OCS and BSA respectively (p>0.05). In conclusions, TCM supplemented with either FBS, OCS or BSA as protein and FSH as hormone may be used as medium for IVM of oocytes of indigenous zebu cows.

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Section: Experiments 3 Determination Of An Effective Protein Supplemementioning

confidence: 87%

Determination of an Effective Media and Its Hormone and Protein Supplemenntation for in Vitro Maturation of Oocytes of Indigenous Zebu Cows

Rahman¹,

Morshed²,

Juyeana³

et al. 2018

Bangl. J. Vet. Med.

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Artificial Meiotic Arrest and Parthenogenetic Activation of Buffalo Oocytes Using Cycloheximide

2007

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Summary Maintaining oocytes at germinal vesicle (GV) stage without damaging their quality would allow synchronization of maturation and homogenization of the oocyte population. Activation of the oocyte is very important for a number of oocyte or embryo related technologies including intracytoplasmic sperm injection and cloning by nuclear transfer. This work will focus on induction of meiotic inhibition and oocyte activation by cycloheximide (CHX), in buffalo. For this purpose 2 experiments were conducted. In experiment I, buffalo cumulus-oocyte complexes (COCs) were cultured in 100 ml droplets of tissue culture medium 199 enriched with 10% v/v fetal calf serum, FSH and LH (0, 05 iu/ml, PERGOVET 75, Serono, Rome, Italy) and 50 mg/ml gentamycin for 24-26 h at 38.5°C in 5% CO 2 and humidified air for 24 h in the presence of 2 mg/ml CHX. COCs were significantly blocked at the GV stage (20.85% vs. 82.72% in control). Reversibility of the CHX effect was assessed by culturing COCs an additional 24 h in CHX-free culture medium. About 65.0% of treated oocytes (control 80.1%) resumed meiosis and progressed to the MII stage. In experiment II, buffalo oocytes were activated by 10 mM calcium ionophore A23187 for 5 min followed by 10 mg/ml CHX for 3 h. Control oocytes were exposed to conventional in vitro fertilization (IVF) in BO medium. After 48 h of culture, significantly more oocytes (pϽ0.05) were cleaved in the IVF group than in treated group and spontaneous activated one. In conclusion, CHX can be used to block spontaneous resumption of meiosis in buffalo oocytes and its effect is reversible. Also, CHX in combination with calcium ionophore have the ability to induce parthenogenetic activation in buffalo oocytes.

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Influence of serum source on buffalo in vitro embryonic development with reference to heat inactivation

Cited by 2 publications

References 26 publications

Determination of an Effective Media and Its Hormone and Protein Supplemenntation for in Vitro Maturation of Oocytes of Indigenous Zebu Cows

Determination of an Effective Media and Its Hormone and Protein Supplemenntation for in Vitro Maturation of Oocytes of Indigenous Zebu Cows

Artificial Meiotic Arrest and Parthenogenetic Activation of Buffalo Oocytes Using Cycloheximide

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