Comparison of Culture Media for In Vitro Maturation of Oocytes of Indigenous Zebu Cows in Bangladesh

Joydev Kumar Singha,Mohammad Musharraf Uddin Bhuiyan,Mohammad Moshiur Rahman,Farida Yeasmin Bari 한국수정란이식학회 2015 한국동물생명공학회지 Vol.30 No.4

The objectives of the present study were to select an effective basic medium including its hormone and protein supplementation for IVM of oocytes of indigenous zebu cows. The ovaries of cows were collected from slaughter house and the follicular fluid was aspirated from 2 to 8 mm diameter follicles. The COCs with more than 3 cumulus cell layers and homogenous cytoplasm were selected for maturation. The oocytes were matured in media for 24 hrs at 39℃ with 5% CO2 in humidified air. The maturation of oocytes was evaluated by examining the presence of first polar body under microscope. An efficient basic medium was determined after culturing COCs in either TCM 199 or SOF medium in Experiment 1. An efficient hormone supplementation was determined after culturing COCs in either FSH or gonadotrophin supplemented TCM 199 in Experiment 2. An efficient protein supplementation was determined after culturing COCs in either FBS or Oestrous cow serum (OCS) supplemented TCM 199 in Experiment 3. The oocyte recovery rate per ovary was 3.35. The overall rate of IVM was 74.6%. The maturation rate was 75.5±3.9 and 62.2±20.2% in TCM and SOF medium, respectively (P>0.05). The maturation rate of oocytes was significantly higher (76.6±13.2%) in FSH supplemented medium than gonadotrphin supplemented counterpart (69.7±10.8%) (P<0.05). The maturation rates of oocytes were 81.7±12.9 and 85.7±12.7% in medium supplemented with FBS and OCS, respectively (P>0.05). In conclusions, both TCM 199 and SOF supplemented with either FBS or OCS, and FSH may be used as medium for IVM of indigenous zebu oocytes in Bangladesh.

Production of cloned sei whale (Balaenoptera borealis) embryos by interspecies somatic cell nuclear transfer using enucleated pig oocytes

이은송,Mohammad Musharraf Uddin Bhuiyan,Hiroyuki Watanabe,Kohji Matsuoka,Yoshihiro Fujise,Hajime Ishikawa,Yutaka Fukui 대한수의학회 2009 JOURNAL OF VETERINARY SCIENCE Vol.10 No.4

In this study, we examined the feasibility of using subzonal cell injection with electrofusion for interspecies somatic cell nuclear transfer (iSCNT) to produce sei whale embryos and to improve their developmental capacity by investigating the effect of osmolarity and macromolecules in the culture medium on the in vitro developmental capacity. Hybrid embryos produced by the electrofusion of fetal whale fibroblasts with enucleated porcine oocytes were cultured in modified porcine zygote medium-3 to examine the effects of osmolarity and fetal serum on their in vitro developmental capacity. More than 66% of the whale somatic cells successfully fused with the porcine oocytes following electrofusion. A portion (60∼81%) of the iSCNT whale embryos developed to the two- to four-cell stages, but no embryos were able to reach the blastocyst stage. This developmental arrest was not overcome by increasing the osmolarity of the medium to 360 mOsm or by the addition of fetal bovine or fetal whale serum. Our results demonstrate that sei whale-porcine hybrid embryos may be produced by SCNT using subzonal injection and electrofusion. The pig oocytes partly supported the remodeling and reprogramming of the sei whale somatic cell nuclei, but they were unable to support the development of iSCNT whale embryos to the blastocyst stage.

A UNIFIED PRESENTATION OF BETA, GAUSS AND CONFLUENT HYPERGEOMETRIC FUNCTIONS

MOHD GHAYASUDDIN,MUSHARRAF ALI,Waseem A. Khan 장전수학회 2022 Advanced Studies in Contemporary Mathematics Vol.32 No.1

In this article, we propose a new extension of beta function by making use of the Bessel-Struve kernel function. Here, first we derive some fundamental properties of this function and then we present a new extension of beta distribution in terms of our proposed beta function. Furthermore, by using the definition of our new beta function, we introduce and investigate a new generalization of Gauss and con uent hypergeometric functions.

In vitro maturation and fertilization of prepubertal and pubertal black Bengal goat oocytes

Momena Khatun,Mohammad Musharraf Uddin Bhuiyan,Jalal Uddin Ahmed,Aminul Haque,Mohammed Shamsuddin,Mohammad Bozlur Rahman 대한수의학회 2011 Journal of Veterinary Science Vol.12 No.1

Oocytes retrieval, in vitro maturation (IVM) and fertilization (IVF) efficiency are inevitable steps towards in vitro production of embryos. In the present study, these parameters were investigated in the ovaries of prepubertal (n = 31) and pubertal (n = 61) black Bengal goats obtained from a slaughterhouse. Nuclear maturation was evaluated upon aspiration and following IVM in TCM-199 (Earle’s salt with L-glutamine and sodium bicarbonate) for 27 h at 39oC under 5% CO2 in humidified air. The oocytes retrieval and efficiency (mean ± SD) per prepubertal and pubertal goats were 5.2 ± 0.6 and 6.8 ± 0.6, and 77.3 ± 0.1% and 80.5 ± 0.6%, respectively. Anaphase I - telophase I stages differed significantly (7.3 ± 0.8 vs. 2.6 ± 0.2, p < 0.05) between the two groups of goats. After IVM, the percentages of metaphase II were significantly higher (66.3 vs. 60.3, p < 0.05) in pubertal goats than in their prepubertal counterparts. The percentages of normal in vitro fertilization (IVF) in Fert-Tyrode’s albumin lactate pyruvate of pubertal goat oocytes did not differ between Percoll and swim-up sperm separation methods (36.7 ± 0.9% vs. 32.7 ± 1.3%, p > 0.05). Furthermore, sperm capacitation by heparin alone or in combination with ionomycin did not lead to a significant increase in the normal fertilization rate (34.8 ± 1.7 vs. 32.2 ± 1.5%, respectively) in the oocytes of pubertal goats. In conclusion, the ovaries of pubertal black Bengal goats obtained from the slaughterhouse could be used for in vitro embryo production. However, further optimization of the IVM and IVF techniques are necessary for satisfactory in vitro embryo production.

Culture Conditions for In Vitro Maturation of Abattoir Derived Oocytes of Native Zebu Cows of Bangladesh

SM Niyaz Morshed,Mohammad Musharraf Uddin Bhuiyan,Mohammad Moshiur Rahman,Joydev Kumer Singha,Nasrin Sultana Juyena 사단법인 한국동물생명공학회 2014 한국동물생명공학회지 Vol.29 No.3

The objectives of the study were to determine an effective culture dish, culture duration and protein supplementationin medium for in vitro maturation (IVM) of oocytes of native zebu cows in Bangladesh. The ovaries of cows werecollected from local slaughterhouse followed by aspiration of follicular fluid. The cumulus-oocyte-complexes (COCs)with more than 3 compact cumulus cell layers were cultured in tissue culture medium (TCM) 199 for maturation. The maturation of oocytes was determined by observing polar body under microscope. To determine an effectiveculture dish, 130 COCs derived from 48 ovaries in a well of 4-well dish and 102 COCs derived from 36 ovariesin drops covered with mineral oil within 35 mm petri dish were cultured for 24 hours. The rate of maturation ofoocytes did not vary between 4-well dish (51.3 ± 15.0 %) and drops in petri dish (52.4 ± 11.6 %). To determine theeffective culture duration, 185 COCs derived from 62 ovaries were cultured in drops for 18, 21, 24 and 27 hours. The rate of maturation of occytes ranged from 51.9 ± 9.4 % (18 hours) to 59.0 ± 17.1 % (27 hours) and the differencein maturation rate among different culture durations was not significant (P>0.05). To determine an effective proteinsupplementation, 63 oocytes from 19 ovaries were cultured separately in TCM 199 supplemented with either fetalbovine serum (FBS) or bovine serum albumin (BSA). The rate of maturation was significantly (P<0.01) higher inmedium supplemented with FBS (55.63 ± 16.19 %) than that of BSA (14.82 ± 9.36 %). In conclusion, COCs of nativezebu cows can be cultured for IVM either in 4-well culture dish or droplets in petri dish for 18 to 27 hours in mediumsupplemented with FBS.

Parthenogenetic Activation of Black Bengal Goat Oocytes

Aminul Haque,Mohammad Musharraf Uddin Bhuiyan,Momena Khatun,Mohammed Shamsuddin 사단법인 한국동물생명공학회 2011 한국동물생명공학회지 Vol.26 No.2

In vitro maturation and activation of oocytes are primary steps towards biotechnological manipulation in embryology. The objectives of the present study were to determine the oocyte recovery rate per ovary, in vitro maturation rates of oocytes and rates of parthenogenetically activation of matured oocytes in Black Bengal goats. All visible follicles were aspirated to recover follicular fluid from individual ovaries (number of ovaries = 456). The immature cumulus oocyte complexes (COCs; n = 1289) were cultured in tissue culture medium (TCM)-199 supplemented with 10% (v/v) fetal bovine serum (FBS) for 27 hours at 39℃ with 5% CO_2 in humidified air. The matured oocytes (n =248) were activated with 5 μM ionomycin for 5 minutes followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) for 4 hours. After activation, oocytes were cultured for another 14 hours in TCM-199 supplemented with bovine serum albumin (BSA) at 39℃ with 5% CO_2 in humidified air. The pronucleus formation in activated oocytes was determined by staining with 1% orcein (whole mount technique). Matured oocytes (n = 176) without activation stimuli were used as control. The mean number of oocytes recovered per ovary was 3.5 ± 0.5. The proportion of oocytes matured in vitro, confirmed by the presence of first polar body, was 42.1 ± 4.7%. Parthenogenetic activation,evidenced by formation of pronucleus, occurred in 37.2 ± 15.8% of matured oocytes. No pronucleus formation was observed in control oocytes. In conclusion, a combination of ionomycin and 6-DMAP induces activation in one third of Black Bengal goats’ oocytes

Comparison of Culture Media for In Vitro Maturation of Oocytes of Indigenous Zebu Cows in Bangladesh

Joydev Kumar Singha,Mohammad Musharraf Uddin Bhuiyan,Mohammad Moshiur Rahman,Farida Yeasmin Bari 사단법인 한국동물생명공학회 2015 한국동물생명공학회지 Vol.30 No.4

The objectives of the present study were to select an effective basic medium including its hormone and protein supplementation for IVM of oocytes of indigenous zebu cows. The ovaries of cows were collected from slaughter house and the follicular fluid was aspirated from 2 to 8 mm diameter follicles. The COCs with more than 3 cumulus cell layers and homogenous cytoplasm were selected for maturation. The oocytes were matured in media for 24 hrs at 39℃ with 5% CO2 in humidified air. The maturation of oocytes was evaluated by examining the presence of first polar body under microscope. An efficient basic medium was determined after culturing COCs in either TCM 199 or SOF medium in Experiment 1. An efficient hormone supplementation was determined after culturing COCs in either FSH or gonadotrophin supplemented TCM 199 in Experiment 2. An efficient protein supplementation was determined after culturing COCs in either FBS or Oestrous cow serum (OCS) supplemented TCM 199 in Experiment 3. The oocyte recovery rate per ovary was 3.35. The overall rate of IVM was 74.6%. The maturation rate was 75.5±3.9 and 62.2±20.2% in TCM and SOF medium, respectively (P>0.05). The maturation rate of oocytes was significantly higher (76.6±13.2%) in FSH supplemented medium than gonadotrphin supplemented counterpart (69.7±10.8%) (P<0.05). The maturation rates of oocytes were 81.7±12.9 and 85.7±12.7% in medium supplemented with FBS and OCS, respectively (P>0.05). In conclusions, both TCM 199 and SOF supplemented with either FBS or OCS, and FSH may be used as medium for IVM of indigenous zebu oocytes in Bangladesh.

Parthenogenetic Activation of Black Bengal Goat Oocytes

Haque, Aminul,Bhuiyan, Mohammad Musharraf Uddin,Khatun, Momena,Shamsuddin, Mohammed 韓國受精卵移植學會 2011 한국동물생명공학회지 Vol.26 No.2

In vitro maturation and activation of oocytes are primary steps towards biotechnological manipulation in embryology. The objectives of the present study were to determine the oocyte recovery rate per ovary, in vitro maturation rates of oocytes and rates of parthenogenetically activation of matured oocytes in Black Bengal goats. All visible follicles were aspirated to recover follicular fluid from individual ovaries (number of ovaries = 456). The immature cumulus oocyte complexes (COCs; n = 1289) were cultured in tissue culture medium (TCM)-199 supplemented with 10% (v/v) fetal bovine serum (FBS) for 27 hours at with 5% in humidified air. The matured oocytes (n = 248) were activated with 5 ionomycin for 5 minutes followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) for 4 hours. After activation, oocytes were cultured for another 14 hours in TCM-199 supplemented with bovine serum albumin (BSA) at with 5% in humidified air. The pronucleus formation in activated oocytes was determined by staining with 1% orcein (whole mount technique). Matured oocytes (n = 176) without activation stimuli were used as control. The mean number of oocytes recovered per ovary was . The proportion of oocytes matured in vitro, confirmed by the presence of first polar body, was . Parthenogenetic activation, evidenced by formation of pronucleus, occurred in of matured oocytes. No pronucleus formation was observed in control oocytes. In conclusion, a combination of ionomycin and 6-DMAP induces activation in one third of Black Bengal goats' oocytes.

Comparison between Two Cryo-devices for Vitrification of Immature Oocytes of Indigenous Zebu Cows in Bangladesh

Sk Mohiuddin Choudhury,Mohammad Musharraf Uddin Bhuiyan,Mohammad Moshiur Rahman,Md. Masudur Rahman,Md. Newaz Sharif,Jayonta Bhattacharjee,Farida Yeasmin Bari,Nasrin Sultana Juyena 사단법인 한국동물생명공학회 2017 한국동물생명공학회지 Vol.32 No.4

Cryopreservation of oocytes by vitrification technique may contribute a lot in the field of reproductive biotechnology. The objectives of the present study were to evaluate the effectiveness of two cryo-devices for vitrification of immature oocytes of indigenous zebu cows. Slaughter house derived immature cumulus-oocyte-complexes (COCs) of cows were vitrified using 15% dimethyl sulphoxide (DMSO) as cryoprotective agent (CPA) with 0.5 mol sucrose in TCM 199 supplemented with 20% FBS. Vitrification of COCs was completed after immediate plunging of COCs loaded cryotop or French mini straw into the liquid nitrogen (LN2). Then the COCs containing cryotop or French mini straws were warmed in 0.25 mol sucrose and 20% FBS supplemented TCM 199 followed by in vitro culture in 50 μl droplets of bicarbonate buffered TCM 199 supplemented with 10% FBS, pyruvate, FSH and oestradiol for 24 hrs at 39°C with 5% CO2 in humidified air. After maturation culture, oocytes were denuded and examined under inverted microscope for presence of polar body as the indication of maturation. Denuded oocytes were also stained by whole mount technique using 1% orcein to examine the maturation by presence of MII chromosomes. The in vitro maturation rate was significantly (p<0.05) higher in oocytes vitrified and warmed using crytop (47.1±6.9%) than that of French mini straw (15.9±12.5%). Moreover, in vitro maturation rate was significantly (p<0.05) highe r in control oocytes (not vitrified) (84.5±14.2%) than that of vitrified oocytes. In conclusion, cryotop is better than French mini straw as cryo-device for vitrification of bovine immature oocytes

Stereoselective Microbial Hydroxylation of Progestin, Norethisterone by Using Aspergillus niger and Penicillium citrinum

Azizuddin,Muhammad Iqbal,Syed Ghulam Musharraf,Saleem Shahzad 한국생약학회 2020 Natural Product Sciences Vol.26 No.4

Microbial transformation of a potent progestin, norethisterone (17β-hydroxy-19-nor-17α-pregn-4-en-20-yn-3-one) (1) was carried out by using two filamentous fungi Aspergillus niger and Penicillium citrinum. Biotransformation of 1 with A. niger yielded a hydroxylated transformed product 10β,17β-diydroxy-19-nor-17α-pregn-4-en-20-yn-3-one (2) whereas 11β,17β-diydroxy-19-nor-17α-pregn-4-en-20-yn-3-one (3) was obtained through microbial transformation of 1 by P. citrinum. It is the first report of their production from 1 by using A. niger and P. citrinum with complete 1H- and 13C-NMR assignment. The structures of both metabolites were characterized by various spectroscopic techniques and reported data.