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Journal : Jurnal Veteriner

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.
Korelasi Antara Lama Kebuntingan, Bobot Lahir dan Jenis Kelamin Pedet Hasil Inseminasi Buatan pada Sapi Bali Gatot Prasojo; Iis Arifiantini; Kusdiantoro Mohamad
Jurnal Veteriner Vol 11 No 1 (2010)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Purpose of the research was to study the correlation between the gestation period with sex and birthweight of new born calves as the result of artificial insemination (AI) in bali cattle. Data were collecteddirectly from AI technicians in some districts in the province of bali from 1997 to 2003. A total, of 10759inseminated cows and 799 of which were with completed data of, date of birth, birth weight and sex ofcalves. The data were collected from 3 districts, involving Gianyar (205 samples), Badung (221 samples)and Bangli (373 samples). Result showed that the gestation period of bali cows were 284.4±5.7 days andbirth weight of 18.4±1.6 kg. Sex ratios (male : female) of bali calves resulted from AI in 1997 to 2003 were1.2:1. There were positive correlation between gestation period and birth weight (P>0.01) as well asbetween gestation period and sex of calves with a high correlation coefficient (0.075) which proved thatmale calves had longer gestation period than females.
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.
Pertumbuhan dan Sekresi Protein Hasil Kultur Primer Sel-Sel Serebrum Anak Tikus (IN VITRO GROWTH AND PROTEIN SECRETION OF NEWBORN RAT CEREBRAL PRIMARY CELLS CULTURE) Ita Djuwita; Vivit Riyacumala; Kusdiantoro Mohamad; Wahono Esthi Prasetyaningtijas; Nurhidayat .
Jurnal Veteriner Vol 13 No 2 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

The objective of this study was to identify the proliferation and differentiation of mice cerebrum cellsand its protein product. Research has been conducted on in vitro growth of three days old rat cerebrum cellsin Dulbecco’s Modified Eagle Medium (DMEM) containing 10% Amino Acid Non Essential (AANE), 10%Fetal Bovine Serum (FBS), 3mM NaHCO3, 50 mg/ml gentamycin, and supplemented with and without 5mg/ml insulin, 10 mg/ml transferin, 5 mg/ml selenium (ITS). Culture was done in 5% CO2, then incubatedat 37oC until 90% confluent. Identification were done on cell proliferation and population doubling time(PDT), number of neuron and glial cells, length of axon and dendrit, and protein secretion. The number andlength of neuronal cells were calculated by using hemocytometer and eyepiece micrometer, respectivelly.Protein secreted into culture medium, designed as conditioned medium (CM) was analysized using sodiumdodecyl sulfate–polyacrilamide gel electrophoresis (SDS-PAGE) method. Quantitative data were analyzedusing statistical T-test on Minitab program. In vitro culture of rat cerebrum cells showed two types of cellsincluding nerve cells of bipolar and multipolar neurons and glial cells including astrocyte (the fibrous andprotoplasmic), oligodendrocyte, and microglia. Supplementation of ITS into the culture medium increasedthe cell proliferation rate (P<0.05) with lower PDT, and quantitatively, the 30 kDa secreted protein asindicated by the higher intensity of the protein band.
Perbedaan Morfologi dan Ekspresi Dazl dan Vasa pada Sel Germinal Fetus dan Anak Mencit Jantan Wahono Esthi Prasetyaningtyas; Ni Wayan Kurniani Karja; Mokhamad Fahrudin; Kusdiantoro Mohamad; Srihadi Agungpriyono
Jurnal Veteriner Vol 24 No 1 (2023)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.19087/jveteriner.2023.24.1.49

Abstract

Sel germinal merupakan salah satu sumber sel yang masih bersifat totipotensi dan berperan dalam pembentukan organisme baru. Morfologi dan ekspresi protein pada sel germinal bersifat dinamis bergantung pada umur dan tahap perkembangan. Penelitian ini bertujuan untuk mengamati perubahan morfologi dan ekspresi protein sebagai marka sel germinal jantan pada fetus umur 13,5 hari pascakawin (days post coital/ dpc) dan anak mencit umur lima hari pascalahir. Hasil Rigi kelamin dan testis diisolasi dari mencit umur 13,5 dpc dan 5 hari. Jaringan kemudian dipreparasi histologi rutin, dan diwarnai dengan pewarnaan hematoksilineosin (HE), sedangkan untuk mengidentifikasi keberadaan protein Dazl, Vasa dan Oct4, jaringan diwarnai dengan pewarnaan imunohistokimia menunjukkan morfologi sel germinal jantan pada fetus mencit umur 13,5 dpc dan anak mencit umur lima hari pascalahir sama-sama berbentuk bulat oval. Namun, sel germinal jantan pada mencit umur lima hari pascalahir berukuran lebih besar, jumlah yang lebih sedikit dan terletak jauh dari membran basal. Pada sel germinal jantan umur 13,5 dpc menunjukkan positif lemah terhadap antibodi Oct 4 dan DAZL serta positif kuat terhadap antibodi Vasa. Pada umur lima hari, sel germinal jantan menunjukkan positif kuat terhadap antibodi Oct-4 dan DAZL, serta positif lemah terhadap antibodi Vasa. Simpulan dari penelitian ini adalah morfologi dan ekspresi marka sel germinal dipengaruhi oleh tahapan pertumbuhan dan perkembangan sel-sel germinal. Vasa dapat digunakan sebagai marka untuk sel germinal umur 13,5 dpc dan DAZL sebagai marka untuk sel germinal umur lima hari pascalahir.