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Ita Djuwita
Departemen Anatomi Fisiologi dan Farmakologi, Fakultas Kedokteran Hewan, IPB, Bogor 16680, Indonesia
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Earth & Planetary Sciences Education Immunology & microbiology Medicine & Pharmacology Veterinary

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Journal : Jurnal Veteriner

 1 
Vitrifikasi Blastosis Mencit dengan Metode Kriolupv ?O I Wayan Batan; I Ketut Suatha; Wahono Esti PrasetyoningtyaserB; Nining Handayani; Ita Djuwita; Arief Boediono
Jurnal Veteriner Vol 10 No 4 (2009)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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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 Ita Djuwita; Faralinda Sari; Kusdiantoro Mohamad
Jurnal Veteriner Vol 11 No 3 (2010)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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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.
Peranan Zona Pelusida Sebagai Barier Terhadap Cemaran Escherichia coli K99 (THE ROLE OF ZONA PELLUCIADA AS A BARRIER OF E COLI K99 CONTAMINATION) I Wayan Batan; Bibiana Widiati Lay; Ita Djuwita; Supar .; Arief Boediono
Jurnal Veteriner Vol 14 No 2 (2013)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

The aim of the study was to investigate the role of zona pelucida (ZP) as a barrier of embryo againstE.coli K99 contamination.   The complete randomized design was used in this study.  The embryos weregiven treatment as a follow : 1) embryos without ZP were contaminated with E.coli K99;  2) embryo withintact ZP were contaminated with E.coli K99;  and  3) embryos  with intact ZP were not contaminated withE.coli K99, as a control.  In each treatment there was 15 replication and in each replication there was oneembryo.  The embryos were incubated in incubator at 37°C and 5% CO2 atmosphere.  The embryos wereobserved every six hours in 24 hours using inverted microscope.  The result showed that embryos withintact ZP could develop in culture contaminated with  E.coli K99, while embryos without ZP becomedegenerated.  The viability of intact embryos was 75% and the embryos without ZP were 65%.  Embryosculture in contaminated medium could develop from eight cells embryo into morulla stage of embryo,compact morulla, and blastocyst. E.coli K99 contamination could inhibit embryo development.  In conclusion,ZP could protect embryo against E.coli K99 contamination.
Aktivasi Oosit Menggunakan Strontium Klorida setelah Injeksi dengan Spermatozoa Domba Hasil Pengeringbekuan (OOCYTE ACTIVATION USING STRONTIUM CHLORIDE FOLLOWING INJECTION OF FREEZE-DRIED RAM SPERMATOZOA) Takdir Saili; Ita Djuwita; Mohamad Agus Setiadi; Srihadi Agungpriyono; Arief Boediono
Jurnal Veteriner Vol 13 No 3 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

One of the factors that inhibit the formation of male pronuclei following injection of freeze-dried ramspermatozoa was the absence of artificial activation during oocyte incubation after the injection. Therefore,in this experiment the ability of strontium chloride (SrCl2) to improve oocyte activation followingintracytoplasmic sperm injection (ICSI) was evaluated. Aceto lacmoid staining was used to assessdecondensation and pronucleus formation following ICSI. Results of this experiment revealed that freezedriedspermatozoa had the ability to decondense and to form 1PN following injection into oocytes evenwithout artificial activation, but failed to form 2PN. However, 40% of 2PN oocytes were obtained when theinjected oocytes was first incubated for 20 minutes in medium containing 50 mM strontium chloride thensubsequently incubated for 10 hours in medium without strontium. On the contrary, the 2PN oocytes werenot observed either in injected oocyte neither without artificial activation nor in non-injected oocytes withartificial activation. In conclusion, freeze-dried ram spermatozoa were able to decondense and to support2PN formation following ICSI and artificial activation using strontium.
Kebuntingan Hasil Transfer Blastosis Mencit yang Dibekukan dengan Metode Vitrifikasi Kriolup I Wayan Batan; I Ketut Suatha; Ita Djuwita; Nining Handhayani; Wahono Esti Prasetyaningtyas; Ketut Adnyane Mudite; Bibiana W Lay; Supar -; Arief Boediono
Jurnal Veteriner Vol 12, No 3 (2011)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

The aim of the study was to assess the viability of vitrified embryo using cryoloop as a carrier ofembryo. The blastocyst stage embryos were collected from superovulated mice. Embryos were frozenusing vitrification method and vitrified embryos were loaded on copper filament cryoloop before dipped inliquid nitrogen. The viability of vitrified embryos was assess in vitro by medium cultered and in vivo bytransfered them to recipient mice. The result shows the viability of vitrified embryos was 85,7% after 24hours cultured and the embryos were born from two pregnant recipient mice out of nine (22%) or fouroffspring out of 63 trasfered embryos (6%). In conclusion, vitrified blatocyst stage embryos using cryoloopas a carrier could keep the viability of the embryos and they could be transfered to the recipient mice andwere born normally.
Black Seed (Nigella sativa) Extract Induce in vitro Proliferation and Differentiation of Rat Pancreatic and Bone Cells (EKSTRAK JINTAN HITAM (Nigela sativa) MENGINDUKSI PROLIFERASI DAN DIFERENSIASI SEL PANKREAS DAN SEL TULANG TIKUS SECARA IN VITRO) Wahono Esthi Prasetyaningtyas; Deny Putra Romadhon; Fitri Susana; Ita Djuwita; Kusdiantoro Mohamad
Jurnal Veteriner Vol 17 No 3 (2016)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Black seed (Nigella sativa), a medicinal plant, widely used for treating various diseases, includingdiabetes mellitus and osteoporosis. This study examined the proliferation and differentiation of pancreaticand bone cells of rat cultured in vitro in medium supplemented with N. sativa extracts (NS). Pancreaticand bone cells were isolated from five days old rat and cultured in Dulbecco modified eagle mediumsupplemented without NS (0%, as control), and with NS (0.05% and 0.5%, as treatment groups) in 5% CO2incubator at 37oC for seven days and observed for cell population doubling time (PDT); proportion anddiameter of Langerhans islets, osteoblast, and osteocyte; and proportion of Langerhans islets containingb cell expressing insulin secretion. The pancreatic b cells were observed using dithizone staining, while thebone cells using alizarin red staining. The result showed that supplementation of NS significantly (p<0.05)decreased the PDT of pancreatic and bone cells, increased the proportion and diameter of Langerhansislets, increased the proportion of expression the b cell producing insulin, and increased the diameter ofosteoblast. In conclusion, the supplementation of NS in culture medium improved the proliferation anddifferentiation of pancreatic and bone cells in vitro.
Kajian In vitro Aktivitas Sel-Sel Trofoblas Blastosis Mencit Aging dan Pengaruhnya terhadap Kegagalan Implantasi Ita Djuwita; Roza Helmita; Adi Winarto; Wahyudin -
Jurnal Veteriner Vol 10 No 1 (2009)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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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.
CARBOHYDRATES OF CHANGES DURING THE FOLLICULAR DEVELOPMENT IN THE OVARY OF THE MOUSE DEER, TRAGULUS JAVANICUS Hamny -; Srihadi Agungpriyono; Ita Djuwita; Chairun Nisa; Wahono Esthi Prasetyaningtyas; Erdiansyah Rahmi
Jurnal Veteriner Vol 9 No 1 (2008)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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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.
Co-Authors Adi Winarto AL-AZHAR AL-AZHAR Ari Purbayanto Arief Boediono Bambang Purwantara Bibiana W Lay BIBIANA W LAY Chairun Nisa Clara M. Kusharto Deny Putra Romadhon Erdiansyah Rahmi Etty Riani Faralinda Sari Fitri Susana Hamny Sofyan Harald Asmus Heru Setijanto I Ketut Suatha I Wayan Batan Iman Supriatna KARTINI ERIANI Katrin Roosita Ketut Adnyane Mudite Koeswinarning Sigit Kusdiantoro Mohamad M Agus Setiadi Mohamad Fakhrudin Mokhamad Fahrudin Muhamad Rizal Martua Damanik N adiarti Nining Handayani Nining Handhayani Nurhidayat - Rimbawan R Roza Helmita SONY HERU SUMARSONO Srihadi Agungpriyono Sugeng Budiharsono Supar - Supar . TAKDIR SAILI Tutik Wrediati Wahono Esthi Prasetyaningtyas Wahono Esthi Prasetyaningtyas Wahono Esti Prasetyaningtyas Wahono Esti PrasetyoningtyaserB Wahyudin - Yohan Rusiyanto Yuhara Sukra