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Number and Quality of Oocytes Collected from Heterotopic Autografted Mice Ovary after PMSG Induction

HAYATI Journal of Biosciences Vol 18, No 4 (2011): December 2011
Publisher : Bogor Agricultural University, Indonesia

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Abstract

Heterotopic grafting sites can be useful in producing oocytes for in vitro Fertilization, therefore, maximising the oocyte yield from the graft by gonadotrophin stimulation would be advantageous. The aim of this study was to investigate the number and quality of oocytes collected from heterotopic autografted ovary after Pregnant Mare Serum Gonadothropin (PMSG) induction. Graft recipients were treated either with or without PMSG stimulation 48 hours prior to graft collection. Ovarian tissue from four weeks old mice (DDY strain) were autotransplanted under the kidney capsule of the same ovariectomized mice and the oocytes were collected 21 days after autotransplantation. The results showed that the average number of oocytes collected from autografted ovaries without PMSG induction were 9.0. + 2.8 not significantly different with those received PMSG induction, 10.9 + 5.1. The percentage of matured and fertilized oocytes and the developed embryos from the autografted ovaries without PMSG induction were 52.4, 33.4, and 26.0%, respectively not significantly different with those received PMSG induction, 53.2, 35.1, and 29.9%, respectively. The number of oocytes and the capacity to matured, fertilized and developed were significantly lower (P < 0.05) compared to the superovulated nongrafted (control) ovaries. In conclusion, PMSG induction on the graft recipients did not significantly increase oocytes yield from grafted heterotopic ovaries. The number and quality of oocytes produced from the autografted ovaries were lower than the superovulated nongrafted ovaries, but still can be used for in vitro embryo production after sequential in vitro maturation and fertilization.

Aktivitas NADH- tetrazolium reductase sel sel trofoblas pada blastosis yang mengalami hatching dan gagal hatching

Jurnal Anatomi Indonesia Vol 1, No 2 (2007)
Publisher : Jurnal Anatomi Indonesia

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Abstract

Implantation is themost critical stage in the establishment of pregnancy. Inmammals, it has been estimated thatbetween 25%and 60%of conceptuses are lost before or at the time of implantation. The objectives of this studywere to investigate the activity of themitochondrial NADH-tetrazoliumreductase, the outgrowth and differentiationof trophoblast cells in in vitro culture of hatched and non hatched blastocyst. Blastocysts were collectedfrom mice cornua utery at day-4 of pregnancy and were divided into 3 groups: blastocysts undergo hatchingwithin 24 hours, 48 hours and non hatching. Embryos were cultured in DMEMmediumin 5%CO2 incubator at37°C for 10 days.The trophoblastsmonolayer were processed forGimsa staining and histochemistry analysisof NADH-tetrazolium reductase activity. The outgrowth of trophoblast cells were measured using eyespiecemicrometer. The results showed that the activity of NADH-tetrazolium reductase of 24 h and 48 h hatchedblastocysts showed higher intensity than the non hatched blastocysts (P<0,05). The trophoblast outgrowthdiameter of 24 h hatched blastocystwas significantly higher than the non hatched blastocyst, but not signifcantlydifferent with the 48 h hatched.Morphology examination using light microscope showed that the trophoblastmonolayer of 24 h hatched blastocyst differentiated into cytotrophoblast, syncytioptrophoblast andspongiotrophoblast after 5 days of in vitro cultured. In conclusion, in in vitro sudy the failure of blastocystsimplantationwas due to the impairment ofNADHdehydrogenase activity in complex Imitochondria and the failureof outgrowth and differentiation of the trofoblast cells.

Development of Domestic Cat Embryo Produced by Preserved Sperms

HAYATI Journal of Biosciences Vol 15, No 4 (2008): December 2008
Publisher : Bogor Agricultural University, Indonesia

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Abstract

The ability to mature and fertilize oocytes of endangered species may allow us to sustain genetic and global biodiversity. Epididymis sperms may be the last chance to ensure preservation of genetic materials after injury or death of a valuable animal. Studies have been conducted to determine wether both epididymis sperms and oocytes can be used to produce viable embryos and offspring. The purpose of this study was to determine how long cats sperms contained in epididymis were remain motile and had intact membranes when preserved at 4 oC, and to determine whether such those preserved sperms are able to fertilize oocytes. Epididymis was preserved immediately in phosphate buffer saline at 4 oC for 1, 3, and 6 days. The observation of sperm quality and viability after preservation was performed by vital staining acrosom and Hoechst-Propidium Iodine. Biological functions of sperms were evaluated by in vitro culture technique for fertilization, micro fertilization and embryonic development rate in CR1aa medium. The results showed that average motility of sperms collected from ductus deferens, cauda and corpus epididymis decreased not significantly (P > 0.05) from 0, 1, 3, and 6 days of preservation times (from 83.0%, 80.2%, 79.0%; 80.9%, 75.0%, 75.5%; 52.0%, 63.2%, 55.0% to 34.6%, 34.6%, 33.3%, respectively). The general results showed that sperms from epididymis preserved for 1, 3, and 6 days can be used for IVF. The rate of embryonal cleavage produced by IVF technique using sperms collected from epididymis preserved for 1-, 3- and 6-days were 33.3, 26.7, and 20.0%, respectively and significantly different (P < 0.05) from that of controll (50.0%). In conclusion, sperms contained in epididyimis preserved at 4 oC in PBS (Phospate Buffer Saline) for 1-6 days can be used to IVF and in vitro production of cat embryos. Key words: gamet, preservation, in vitro fertilization

In Vitro Fertilization and Embryo Development

HAYATI Journal of Biosciences Vol 12, No 2 (2005): June 2005
Publisher : Bogor Agricultural University, Indonesia

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Abstract

Experiments were conducted on the morphology, fertilization and embryo development rate of vitrified ovine oocytes matured in vitro. Three vitrification solutions were used for vitrification. PBS supplemented with 1% BSA, 30% ethylene glycol was added by one of three different sucrose concentrations, 1.00 M (VS1), 0.50 M (VS2), and 0.25 M (VS3). The results showed that the percentages of normal vitrified oocytes after warming were 78 and 63% in VS1 and VS2, respectively, which was significantly higher as compared for VS3. The fertilization rates were 59 and 66% in VS1 and VS2, respectively, which were also significantly higher as compared with VS3 (35%). Zygote viability after 18 h was 57; 43; and 40%, for VS1,VS2, and VS3, respectively, which was not significantly different. The incidence of polyspermic penetration increased with increasing sucrose concentration, i.e 23, 11, and 9% in VS1, VS2, and VS3, respectively, as compared with unvitrified oocytes (4%). The cleavage rate of vitrified oocytes in VS1 was 13.2% which was significantly lower (p

Allotransplantasi Testis Mencit Muda sebagai Upaya Preservasi Gonad In Vivo

Jurnal Ilmu Pertanian Indonesia Vol 12, No 1 (2007): Jurnal Ilmu Pertanian Indonesia
Publisher : Institut Pertanian Bogor

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Abstract

Cancer disease are not detected only at adult but also at young age. One therapy for cancer diseases is chemotherapy and radiation, that give a side effect of infertility in the gonad, therefore, it is necessary to preserve the gonad. Sperm collection from adult is easy but not from the young patients.Keyword: transpalantation, testis, mencit muda, spermatogenesis

TAHU MENGHAMBAT KEHILANGAN TULANG LUMBAR TIKUS BETINA OVARIEKTOMI [Tofu Attenuates Lumbar Bone Loss of Ovariectomized Female Rats]

Jurnal Teknologi Dan Industri Pangan Vol 13, No 3 (2002): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

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Abstract

TAHU MENGHAMBAT KEHILANGAN TULANG LUMBAR TIKUS BETINA OVARIEKTOMI   [Tofu Attenuates Lumbar Bone Loss of Ovariectomized Female Rats] Samsu Udayana Nurdin 1) , Deddy Muchtadi 2) , Ita Djuwita 3) , Suyanto Pawiroharsono 4) 1) Department of Agricultural Product Technology, Lampung University, Jln. Sumantri Brojonegoro No. 1Bandar Lampung, 35145;  2) Department of Food Technology and Human Nutrition, Bogor Agriculture University, Jln. Lingkar Kampus, Dramaga Bogor; 16680; 3) Laboratory of Embriology, Veteriner Faculty, Bogor Agriculture University, Jln. Agatis, Dramaga Bogor; 16680; 4) Agency for the Assesment and Application of Technology, Jln. MH. Thamrin no. 8, Jakarta 10340. ABSTRACT   The objectives of this research were to examine the efeects of feed containing soybean tofu and tempeh on lumbar bone density and mass of ovariectomized female rats. Twenty four 17 weeks-old Sprague-Dawley rats were randomly assigned to four group, i.e.: (1) non-ovariectomized rats fed casein based diet (NonOvx), (2) ovariectomized rats fed casein based diet (OvxC), (3) ovariectomized rats fed diet containing soybean tofu (OvxH), and (4) ovariectomized rats fed diet containing soybean tempeh (OvxT); in three block based on their body weight.  The result show that body weight gram of ovariectomized rats was greater than nonovariectomized.  Ovariectomy caused atrophy of the uterus, and resulted in higher serum calcium level.  The lower lumbar vertebrae density of ovariectomized rats was observed and the decrease was prevented by tofu.   Key words : Tofu, tempeh, bone loss,  and ovariectomy

CARBOHYDRATES OF CHANGES DURING THE FOLLICULAR DEVELOPMENT IN THE OVARY OF THE MOUSE DEER, TRAGULUS JAVANICUS

Jurnal Veteriner Vol 9, No 1 (2008)
Publisher : Jurnal Veteriner

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Abstract

The data available on the female reproductive organ of mouse deer (Tragulus javanicus) is still very limited. A study was therefore conducted to investigate the distribution and the concentration of carbohydrate residues during the development of ovary follicles. An ovary at luteal phase was used in this study. Thin sections of the ovary were prepared occording to the standard methods and they were then histochemically stained with flourecnece-labelled lectins such as peanut agglutinin (PNA), Ricinus communis agglutinin (RCA), Concanavalin A (Con A), Winged bean agglutinin (WGA) and Ulex europaeus agglutinin (UEA). The result showed that changes in the distribution and the concentration of carbohydrate occured during the development of the follicle. During the preantral stage, the cytoplasm of oosit contained carbohydrate with the residues of glucosa dan mannosa. Zona pelusida contained carbohydrates with residues of glucosa, mannosa, galactosa dan N-asetylgalactosamine, whereas extracellular matrix contained carbohydrate with the residues of glucosa dan mannosa. In the antral follicle, the cyitoplasm of oocytes contained carbohydarte with the residues of galactosa dan N-asetylgalactosamine, whereas its zona pelusida, extracellular matrix and follicular fluid contained carbohydarte with the residues of fucosa, N-asetylglucosamin and cyalic acid. Diffrences in the types and the distribution pattern of carbohydrates were observed in this study, both in preantral and antral follicles.

Kajian In vitro Aktivitas Sel-Sel Trofoblas Blastosis Mencit Aging dan Pengaruhnya terhadap Kegagalan Implantasi

Jurnal Veteriner Vol 10, No 1 (2009)
Publisher : Jurnal Veteriner

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Abstract

The objectives of this in vitro study were to investigate the hatching rate, the outgrowth diameter andthe activity of mitochondria Nicotiamide Adenin Dinucleotide Dyhidrogenage (NADH)-CoQ reductase ofblastocysts trophoblast cells from aging mice. Blastocysts of aging (age >12 months) and young productive(age 2 months) mice were collected from the cornua utery at day-4 of pregnancy and were cultured inmDMEM medium supplemented with 10% New Born Calf Serum (NBCS), 10% ITS, and 50 ?g/mlgentamicine, in 5% CO2 incubator at 37°C for 10 days. The blastocysts hatching rate and the trophoblastsmonolayer were examined for their diameter outgrowth and the NADH-CoQ reductase activity. The resultsshowed that the hatching rate, the trophoblast outgrowth diameter and the activity of NADH-CoQ reductaseof blastocysts collected from productive mice were significantly higher than those collected from the agingmice (P<0,05). It can be concluded that the impairment of blastocysts implantation especially, in agingmice were caused by the low activity of the NADH-CoQ reductase that play important role in energyproduction needed for the hatching and trophoblast outgrowth.

Vitrifikasi Blastosis Mencit dengan Metode Kriolupv ?O

Jurnal Veteriner Vol 10, No 4 (2009)
Publisher : Jurnal Veteriner

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Abstract

Cryopreservation is an ultra rapid freezing process to preserve tissue or organ. The studywas conducted to identify the effectiveness of cryoloop vitrification method and the viability ofembryos following vitrification. Embryos at blastocyst stage were vitrified by placing them inequilibration medium containing 10% ethylene glycol (EG) and phosphate buffer saline (PBS) wichsupplemented with 20% new born calf serum for 8-10 minutes. The blastocysts were then removedand put in vitrification medium (15% dimethyl sulfoxide, 15% EG, and 0.5M sucrose), and theprocess in the vitrifivcation medium not longer than 25-30 seconds. The blastocysts were immediatelytransferred to the vitrification medium film in the cryoloop and plunged into 100 ml liquid nitrogen.The warming process was done by immersing the cryoloop which carried the vitrified blastocstsinto PBS supplemented with 20% serum and 0.5M sucrose for 1 minute, and then removed to samesolution supplemented with 0.25M sucrose and 0.1M sucrose for 2 minutes respectively. Theblastocysts were washed 4 times in kalium simplex optimized medium (KSOM) and cultured indrops of KSOM in 5% CO2 incubator at 370C. The observations were done every 6 hours for 48hours using inverted microscope ( Olympus IX70 Japan). The viability of embryos was assessed onthe basis of the intact morphology, reexpansion of the blastosul, and the development of embryosinto advance stage. The results showed that 85.71% of vitrified embryos, developed into advancestages and 19% of them hatched. In conclusion the cryoloop can be used to vitrify the embryos.

The Comparison of One and Two Steps Equilibration in Vitrification Process on The Morphology and Viability of Mouse Blastocysts

Jurnal Veteriner Vol 11, No 3 (2010)
Publisher : Jurnal Veteriner

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Abstract

A study was conducted to compare the effect of one and two steps equilibration method of vitrificationon the morphology and viability of mouse blastocysts. Blastocysts were firstly exposed to modified PhosphateBuffered saline (mPBS) containing 1% Bovine Serum Albumin (BSA) proceeded by exposure in mPBSrespectively containing 0.25M sucrose (S) for 2 minutes . Blastocysts were then exposed for 2 minutesrespectively to mPS+0.5M S (one step method) or in mPBS+0.5M S+10% ethylene glycol (EG) (two stepmethod).. Blastocysts were then exposed in mPBS+0.5M S+30% EG for 60 second, loaded into 0.25 mlplastic straw, and exposed immediately in vapor of liquid nitrogen for 10 second before they were and thenplunged into liquid nitrogen. The blastocysts were reconstituted by diluting with mPBS+0.5M S followedby mPBS+0.25M S for each 3 min and washed in mPBS without sucrose. The viability of cells was assessedby fluorescent vital staining, by re-expansion for 24 hours in vitro culture, and by implantation into therecipient oviduct. The percentages of morphologically normal blastocysts following recovery fromvitrification were higher (p<0.05) in one step equilibration than in those of two steps methods (89.6%. vs82.6%). The viability of blastocysts examined under light microscope after staining with biz-benzimidizepropidiumiodine and 24 hours in vitro culture in one step methods (64.0%; 57.8%) were higher (p<0.05)compared with two steps methods (40.0%; 35.6%), respectively. The implantation rate of vitrifiedblastocysts (23.1%) was not significantly different to that of fresh blastocysts (33.4%). These resultsshowed that the one and two step equilibration methods are effective for vitrification and maintaining theviability of the mouse blastocysts.