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Sel Kumulus Sebagai Feeder Layer pada Kultur Stem Cells Embrionic Mencit (CUMULUS CELLS AS FEEDER LAYER IN CULTURE OF MOUSE EMBRYONIC STEM CELLS) Thomas Mata Hine; Arief Boediono; Iman Supriatna; Dondin Sajuthi
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 purpose of this study was to verify the effectiveness of cumulus cells as a feeder layer in supportingthe growth of mouse embryonic stem cells. Embryos at blastocyst stage were exposed in pronase solution,and then incubated in rabbit anti-mouse antibody and guinea pig complement to lyse and separate thetrophoblast cells from the inner cell mass. Inner cell mass subsequently cultured in a petri dish containinga cumulus feeder layer, mouse embryonic fibroblasts, or without a feeder layer, in stem cells medium. Theresulting stem cells colony passaged in trypsin solution, pipetted repeatedly to produce subcolonies orsingle cells, and cultured as before in some new petri dishes. Characterization of stem cells was identifiedby using alkaline phosphatase staining. The results showed the effectiveness of cumulus cells as feederlayer for culture of mouse embryonic stem cells was comparable with mouse embryonic fibroblasts, andboth of them were better than without a feeder layer method. The number of primary colony, cell lines, andcolony growth rate increased 41.30, 8.70, and 54.20%, respectively, while doubling time was shorter 10:21hours compared to the growth without feeder layer method. These results prove that the cumulus feederlayer effectively supports the growth of mouse embryonic stem cells.
MICROVOLUME OF 0.1µL GAMA SLEEVED CRYOLOOPS FOR BLASTOCYST VITRIFICATION OF ASSISTED REPRODUCTIVE TECHNOLOGY PATIENTS Hanoum, Ita Fauzia; Boediono, Arief; Pangestu, Mulyoto; Haryadi, Dwi; Widad, Shofwal; Dasuki, Djaswadi
JURNAL KESEHATAN REPRODUKSI Vol 2, No 1 (2015)
Publisher : Fakultas Kedokteran, Kesehatan Masyarakat dan Keperawatan UGM

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (181.875 KB) | DOI: 10.22146/jkr.7127

Abstract

Ita Fauzia Hanoum1,2, Arief Boediono3, Mulyoto Pangestu4,5, Dwi Haryadi1,Shofwal Widad1,2, Djaswadi Dasuki1,2 ABSTRAK Latar Belakang: Prosedur embrio vitrifikasi menggunakan alat berupa grid, straw atau cryoloop. Gama Sleeved cryoloop dibuat dan dikembangkan di klinik Permata Hati. Untuk itu, dilakukan pengamatan keberhasilan prosedur vitrifikasi menggunakan 0.1µl Gama Sleeved cryoloop.Metode: Vitrifikasi dilakukan pada blastokis dengan kualitas baik yang diperoleh pada hari ke 5 setelah fertilisasi. Inform consent telah disampaikan sebelumnya kepada pasien program bayi tabung di Klinik Permata Hati. Prosedur dilakukan dengan menggunakan media handling (GMOPS Plus; Vitrolife) embrio diinkubasi selama 1 menit; (7.5% EG (v/v); 7.5% DMSO (v/v)) selama 2-3 menit, (15% EG (v/v); 15% DMSO v/v; 10 mg/ml Ficoll; 0.65 M Sucrosa) selama 30 detik pada suhu ruang sebelum kemudian diletakkan di dalam cryoloop, setelah itu secara cepat cryoloop yang berisi embrio dibenamkan ke dalam nitrogen cair. Sebelum dilakukan embryo transfer (ET), embrio dihangatkan dengan cara two step technique (sucrose 0.25M) selama 2 menit dan selama 3 menit (sucrose 0.125M).Hasil: Sejumlah 97 blastokis divitrifikasi dan dihangatkan (67 pasien), dimana 91 blastokis berhasil ditransfer ke rahim ibu (93.8%). Blastokis yang tidak berhasil selamat dari prosedur penghangatan adalah blastokis dengan kerusakan lebih dari 50%. Diperoleh kehamilan klinis 43.3% sedangkan angka implantasi adalah 37.4%. Sampai saat ini, dilaporkan 20 kelahiran (23 bayi) dari program vitrifikasi menggunakan 0.1µl Gama Sleeved cryoloop, sementara 5 kehamilan masih berlangsung. Satu kehamilan dilaporkan gugur pada usia kehamilan yang masih sangat awal, dua keguguran pada usia kehamilan 12 minggu dan satu bayi lahir meninggal karena kelainan kongenital.Kesimpulan: 0.1µl Gama Sleeved cryoloop merupakan pilihan untuk digunakan sebagai alat vitrifikasi blastokis. Data awal yang kami sampaikan dan kelahiran bayi dari program tersebut memberikan harapan untuk kesuksesan program simpan beku embrio di klinik Permata Hati RSUP DR Sardjito Yogyakarta.Kata kunci: kriopreservasi, blastokis, vitrifikasi ABSTRACTBackground: Vitrification has been applied succesfully in human embryo using grid, straw and cryoloop. Gama Sleeved is a home made device develop at Permata Hati. We assessed the survival rate of human blastocyst vitrified in 0.1µl Gama Sleeved cryoloop as device.Method: Excess good grade human D5 embryos were vitrified, upon a detailed informed consent. Embryos were hold in handling media (GMOPS Plus; Vitrolife) for 1 minute; (7.5% EG (v/v); 7.5% DMSO (v/v)) for 2-3 minutes, (15% EG (v/v); 15% DMSO v/v; 10 mg/ml Ficoll; 0.65 M Sucrosa) for 30 seconds at room temperature before inserted in to the loops, then directly plunged into the liquid nitrogen. Prior to ET, embryos were warmed by two step technique in sucrose 0.25M for 2 min and 0.125M sucrosa for 3 min. Embryos were then cultured.Results: Total of 97 vitrified warmed human blastocyst (67 patients) were used and 91 (93.8%) were transferred. Non-transferred blastocyst (6.2%) has more than 50% lyse. The clinical pregnancy rate was 43.9%. The implantation rate was 37.4%. Currently, 20 deliveries of 23 babies born from vitrified blastocyst using 0.1µl Gama Sleeved cryoloop, and another 5 ongoing pregnancy. So far there was 1 early pregnancy loss, 2 miscarriages at 12 weeks pregnancy, and one infant died due to a congenital anomaly.Conclusion: 0.1µl Gama Sleeved cryoloop provides an excellent alternative to existing vitrification devices. These initial data and babies delivered from the program have been promising to a vitrification system in our own ART program.Keywords: cryopreservation, blastocyst, vitrification1Permata Hati Infertility Clinic RSUP DR Sardjito, Yogyakarta2Div Reproductive Endocrinology and Fertility OBGYN Medical Faculty Gadjah Mada University, Yogyakarta3Lab. Anatomi Embriologi FKH, Institut Teknologi Pertanian, Bogor4EPRD- Dept. Obstetrics and Gynecology, Monash University, Monash Medical Center,Victoria, Melbourne5Lab. Reproductive Physiology, Jenderal Soedirman University, Purwokerto Correspondence address: + 62 274 518684; fax + 62 274 553575; email: itafauzia@yahoo.com
KUALITAS, KEMAMPUAN IMPLANTASI DAN VIABILITAS IN-VIVO EMBRIO MENCIT (MUS MUCULUS) GALUR SWISS WEBSTER SETELAH PEMBEKUAN DENGAN METODE VITRIFIKASI Sumarsono, Sony H.; Boediono, Arief; Kusumaningtyas, Hartanti; Madihah, Madihah
Biota : Jurnal Ilmiah Ilmu-Ilmu Hayati Vol 11, No 2 (2006): June 2006
Publisher : Universitas Atma Jaya Yogyakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (364.641 KB) | DOI: 10.24002/biota.v11i2.2618

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Reproductive technologies including in vitro fertilization (IVF), embryo manipulation, gamete and embryo freezing, thawing and embryo transfer were rapidly developed. Vitrification is an embryo freezing technique that is the most developed. In this experiment, we vitrified mouse embryos and then examined the embryos i.e: (i) the quality of the embryos after thawing, (ii) the implantation rate of the embryos and (iii) viability of the embryos in vivo. Morulae and blastocycsts were collected from female mice that were pregnant a day 3,5. The embryos were equilibraten in mPBS +10% etilene glycol. Vitrification was carried out by using VABEDS medium, containing 6-10 embryos that were dropped into a tip of a straw, then frozen in liquid nitrogen for 24 hours. Thawing was carried out by flushing the embryos using mPBS suplemented with 0.5, 0.25, 0.1 and 0 M sucrose. After being incubated in M2 medium at 37oC for 1-2 hours, the recovery embryos were then transferred into the uteri of day 2.5 of pseudopregnat females. The females were then sacrificed at day 16 of gestation and the total implantaion, total life and death fetuses, as well as resorpted embryos, were taken as data. The results showed that vitrification significantly (p<0,05) reduced the quality of the embryos, as well as their implantation rate and the viability of the fetuses, which may be caused by the unoptimal combination of the cryoprotectant in the vitrification medium, temperature and exposure time during vitrification.
Low Concentration of Ethylene Glycol Improved Recovery Rate of Human Spermatozoa After Vitrification (ETILEN GLIKOL KONSENTRASI RENDAH MENINGKATKAN RECOVERY RATE SPERMATOZOA MANUSIA PASCAVITRIFIKASI) Rini Widyastuti; Sony Heru Sumarsono; Arief Boediono; Siti Darodjah Rasad
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

The use of cryoprotectants for the cryopreservation of human spermatozoa, oocytes, zygote, earlycleavage stage of embryos and blastocyst is an integral part of almost every human In Vitro Fertilizationprogram. Moreover, the cryopreservation of these types of cells by direct plunging into liquid nitrogen (-196°C) usually requires a high concentration of cryoprotectant with a consequent of cytotoxic effect. Theaim of this study was to observe the effect of ethylene glycol concentration on the spermatozoa recoveryrate following vitrification process. Earle’s balanced salt solution + 0.25 M sukrosa + 1 % human albumineserum as basic solution supplemented with some different concentrations of etylene glycol (ie: 36.25%;18.25%; 9.12%; 4.56%; 1.14% and 0.57%) were used to evaluate the motility and viability of spermatozoafollowing vitrification. Human’s spermatozoa from ejaculates with progressive motility and viability above50% were used as samples. Samples were mixed with vitrification solution and then loaded into 0.25 mLstraws, equilibrated for 10 minutes at room temperature before plunged into liquid nitrogen. Spermatozoathawing was done at 24 hours after the vitrification. The results showed that, the decrease of spermatozoamotility and viability were observed at the highest (100%, 96.70%, respectively) in the samples that wereadded with vitrification medium contained 36.25% of ethylene glycol. On the other hand, the decrease ofthe spermatozoa motility and viability were found at the lowest (14.11%, 43.81 %, respectively) in thesamples without ethylene glycol supplementation. It can be concluded that the highest spermatozoa recoveryrate was obtained from the vitrification using a low concentration of ethylene glycol.
CONDITIONED MEDIUM DARI KULTUR PRIMER SEL SYARAF MUS MUSCULUS Boediono, Arief; Sandra, Ferry; Puspitasari, Riris L.
Prosiding Seminar Biologi Vol 10, No 2 (2013): Seminar Nasional X Pendidikan Biologi
Publisher : Prodi Pendidikan Biologi FKIP UNS

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Secara  in vitro,  Embryonic Stem Cell  (ESC) dapat diarahkan perkembangannya menjadi sel neuron dan sel glia. Conditionedmedium  dari kultur primer sel syaraf mengandung sejumlah faktor pertumbuhan antara lain  nerve growth factor (NGF), glial derived-neurotrophic factor (GDNF), nestin, dan glial fibrillary acidic protein (GFAP). Dengan melakukan purifikasi protein yang terkandung  di dalam CM, maka diharapkan spektrum protein yang ada menjadi lebih sempit sehingga protein target dapat terdeteksi. Penelitian ini mempelajari kultur primer sel syaraf yang berasal dari hemisfer Mus musculus. Tujuan penelitian adalah untuk mendapatkan CM dari kultur primer sel syaraf  Mus musculus. Medium yang digunakan adalah Dulbecco?s Modified Eagle?s Medium (DMEM) highglucose FBS 10%. Penggantian medium kultur dilakukan setiap 2 hari sekali. Kepadatan sel sekitar 32x103 sel/2 cm2. Setelah hari ke-4 terlihat adanya pertumbuhan neuron bipolar dan neural progenitor cell (NPC). Sel-sel astrosit akan teramati ketika periode kultur diperpanjang. Sel mengalami konfluensi setelah 12 hari kultur. Sel-sel yang tumbuh berguna untuk penjelasan neurogenesis. Kultur primer sel syaraf secara monolayer yang berasal dari hemisfer neonatus mampu mendukung  pertumbuhan sel yang tergolong sebagai neurogenic dan nonneurogenic.  Kata kunci: kultur primer, sel syaraf, conditioned medium, neural progenitor cell, neurogenesis.
Improving the probability of pregnancy in endometriosis cases: a study in patients undergoing in vitro fertilization Moeloek, Farid A.; Pakasi, Trevino A.; Boediono, Arief; Jamaan, Taufik
Medical Journal of Indonesia Vol 21, No 3 (2012): August
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (381.662 KB) | DOI: 10.13181/mji.v21i3.497

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Background: Endometriosis is the most common condition underlying infertility in women. To increase the rate of pregnancy in women with endometriosis, embryo selection is performed during in vitro fertilization. This study aims to prove the effectiveness of embryo selection on the rate of pregnancy in a hospital in Jakarta.Methods: This is a cross sectional clinical epidemiology study, performed on endometriosis patients who visited the hospital between 2007 – 2009. Patients were diagnosed with endometriosis using the laparoscopy technique. Embryo selection was performed by assessing the morphology and cell count.Results: We were able to collect data from 72 subjects who underwent IVF during this research period. One subject was dropped out of the program due to immaturity of the oocyte. Successful fertilization was achieved for 65 subjects, but two of them did not undergo embryo transfer. Out of all the subjects undergoing embryo transfer, 26 subjects successfully became pregnant (36.1%). In severe endometriosis cases, pregnancy was achieved with excellent quality embryos (50%) and good-moderate quality embryos (16.7%); but the probability of failure to become pregnant was found to be the same (50%). In mild-moderate endometriosis cases, the probability of pregnancy with excellent quality embryos was 39% compared to 25% chance with good-moderate quality embryos. Regarding the number of embryos that were transferred, we have found that the probability of pregnancy was 50% higher when 3 embryos were transferred, compared to 1 or 2 transferred embryos.Conclusion: This study shows that embryo quality and the number of transferred embryos are relevant to increase the probability of pregnancy in patients undergoing IVF. But because the probability of not achieving pregnancy is not significantly different, we need to find another marker that is more sensitive to assess the quality of embryo transfer. (Med J Indones. 2012;21:147-51)Keywords: Embryo transfer, endometriosis, oocyte and embryo quality, pregnancy rate
Development of Domestic Cat Embryo Produced by Preserved Sperms KARTINI ERIANI; ARIEF BOEDIONO; ITA DJUWITA; SONY HERU SUMARSONO; AL-AZHAR AL-AZHAR
HAYATI Journal of Biosciences Vol. 15 No. 4 (2008): December 2008
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (90.643 KB) | DOI: 10.4308/hjb.15.4.155

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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
Albumin Telur Sebagai Lem pada Operasi Cangkok Konjungtiva Kartiwa, R. Angga; Enus, Sutarya; Boediono, Arief; Miraprahesti, Retti N.
Majalah Kedokteran Bandung Vol 48, No 4 (2016)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15395/mkb.v48n4.925

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Cangkok konjungtiva sudah lama digunakan pada bidang oftalmologi. Metode yang digunakan saat ini untuk menempelkan cangkok konjungtiva adalah menggunakan teknik jahitan dan lem fibrin. Pada penelitian ini dilakukan uji coba menggunakan lem albumin pada cangkok konjungtiva kelinci sebagai alternatif lain selain menggunakan teknik jahitan dalam penempelan cangkok konjungtiva. Tujuan penelitian adalah membandingkan penyembuhan luka cangkok konjungtiva bulbi antara teknik lem albumin dan jahitan pada mata kelinci. Dilakukan animalexperimental study pada 32 mata (16 ekor kelinci) di PT. Bio Farma (Persero) dan laboratorium Histologi, Fakultas Kedokteran Universitas Padjadjaran dari bulan Maret 2014–Juli 2014, terbagi kelompok teknik lem albumin dan teknik jahitan. Dilakukan pemeriksaan meliputi perbandingan derajat perlekatan cangkok konjungtiva bulbi pada teknik lem albumin dan teknik jahitan yang diamati hari-1 pascabedah serta dilakukan pemeriksaan histologis secara mikroskopik untuk mendapatkan data celah luka yang diamati 10 menit dan hari-7 pascabedah. Analisis data dilakukan dengan Mann-Whitney test for small sample. Hasil penelitian memperlihatkan perlekatan cangkok konjungtiva bulbi secara bermakna lebih kuat pada teknik lem albumin (derajat 4) dibanding dengan teknik jahitan (derajat 2 dan 3) pada hari-1 pascabedah dengan nilai p=0,000 serta terdapat perbedaan celah luka (wound gap) bermakna antara teknik lem albumin (0–0,33 µm) dan jahitan (5,33–14 µm) (p=0,0005)pada cangkok konjungtiva dilihat sepuluh menit pascabedah dan pada hari-7 pascabedah untuk teknik lem albumin (0 µm) dan teknik jahitan (0,33–4 µm) dengan nilai p=0,0005. Simpulan penelitian ini adalah derajat perlekatan jaringan cangkok pada teknik lem albumin lebih baik dibanding dengan jahitan hari-1 pascabedah, sedangkan celah luka lebih kecil pada teknik lem albumin dibanding dengan teknik jahitan pada pengamatan 10 menit dan hari-7 pascabedah. [MKB. 2016;48(4):241–8]Kata kunci: Jahitan, lem albumin, penyembuhan luka konjungtivaEgg Albumin as Adhesive in Conjunctival Graft SurgeryCConjunctival graft has been frequently used in the field of ophthalmology. The frequently used methods to attach a conjunctival graft are suture technique and the use of fibrin glue. This study was to investigate albumin glue as an alternative to suture technique in attaching conjunctival grafts in rabbits. The aim of this study was to compare the conjunctival wound healing between albumin glue and suture technique in rabbit eye as a model. This was an experimental animal study that included 32 eyes (16 rabbits) conducted at PT. Bio Farma (Persero) and the Histology Laboratory, Faculty of Medicine, Universitas Padjadjaran from March 2014 to July 2104. The subjects in this study were divided into albumin glue group and suture technique group. The examinations were comparison of conjunctival graft attachment and histologic microscopic examination to assess the wound gap. Data analysis was performed statistically using Mann-Whitney test for small sample. The statistical analysis results showed that the graft attachment was significantly better when using albumin glue (grade 4) compared to suture (grade 2–3) on day-1 after surgery (p=0.000). The wound gap was smaller using albumin glue (0-0,33 µm versus 5,33-14 µm; p0.0005) 10 minutes after surgery and 0 µm versus 0.33–4 µm, p 0,0005, on day-7 after surgery. In conclusion, graft attachment using albumi n glue is better and the wound gap is smaller when using albumin glue compared to the suture technique. [MKB. 2016;48(4):241–8]Key words: Albumin glue, conjunctival wound healing, suture
STUDY ON SPERM AGGLUTINATION WITH CHARACTERIZATION OF PLASMACOLLECTED FROM EPIDIDYMIS AND EJACULATE IN RAM Muhammad Haviz; Arief Boediono; M Agus Setiadi; Srihadi Agungpriyono; Mokhamad Fahrudin
Jurnal Veteriner Vol 9 No 4 (2008)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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This study was designed to optimalize the use of epididymal or ejaculate sperm and plasma for in vitro fertilization, that sperm agglutination was found at preparation. The rate of sperm agglutination was calculated the head-to-head sperm agglutination that were incubated in Krebs Ringer-(N-(2hydroxyethyl)piperazine-N’-(2-ethenesulfonic acid) or KR-HEPES medium in 38.50C with 5% CO2 at 1, 3, 5 and 7 hours culture in vitro. The rate of head-to-head sperm agglutination were decreased with time treatments. The cauda of sperm agglutination was lower than that caput, corpus epididymal and ejaculate sperm with statistically significant (P<0.01). These result reflected that distribution of anti-agglutinin might be higher in cauda epididymal than that other areas. Number of protein were characterize with SDS-PAGE as follow 11 bands in caput epididymal, 9 bands in corpus epididymal, 2 bands in cauda epididymal and 4 bands in seminal plasma. The higher distribution of protein was found at range 25-40 kDa in epididymal plasma of ram. However, further investigation should be conducted to determine presumptive anti-agglutinin by advance method.
Role of various sugars in improving frozen semen quality of Garut ram Rizal, Muhammad; ., Herdis; Boediono, Arief; Aku, Achmad Selamet; ., Yulnawati
Indonesian Journal of Animal and Veterinary Sciences Vol 11, No 2 (2006)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (86.223 KB) | DOI: 10.14334/jitv.v11i2.516

Abstract

Ram spermatozoa are sensitive to extreme changes in temperature during the freeze-thawed process. The present study was conducted to examine the effects of addition of various sugars in Tris extender on sperm cryosurvival of Garut ram. Semen was collected using an artificial vagina from three mature rams once a week. Immediately after initial evaluation, semen was divided into five parts and diluted with Tris extender (control), Tris extender + 0.4% dextrose, Tris extender + 0.4% raffinose, Tris extender + 0.4% trehalose, and Tris extender + 0.4% sucrose, respectively. Semen was loaded in to 0.25 ml mini straw with the concentration of 200 million or 800 million motile spermatozoa per ml. Semen was equilibrated at 5oC for three hours, then frozen and stored in liquid nitrogen for seven days. Quality of processed-semen including percentages of motile spermatozoa (MS), live spermatozoa (LS), intact acrosome cap (IAC), and intact plasma membrane (IPM) were evaluated after dilution, equilibration, and thawing, respectively. Data were analyzed using completely randomized design with five treatments and six replicates. Means were compared significant difference test at 0.05 significant level. Results of this research showed that there was no significantly difference (P&gt;0.05) between treatments for all sperm quality parameters after dilution and equilibration. Mean percentages of post thawing MS, LS, IAC, and IPM for dextrose (54.00; 68.00; 66.60, and 57.83%), raffinose (50.00; 64.33; 61.80, and 61.75%), trehalose (50.83; 65.67; 61.40 and 57.75%), and sucrose (49.00; 66.80; 58.50 and 58.50%) were significantly (P&lt;0.05) higher than control (40.83; 52.67; 54.60, and 49.40%) respectively. In conclusion, addition of 0.4% dextrose, raffinose, trehalose or sucrose in Tris extender are effective in improving frozen semen quality of Garut ram. Key Words: Sugars, Tris extender, Frozen Semen Quality, Garut Ram
Co-Authors . Alimuddin . Andriyanto . AULANI’AM . Herdis . Yulnawati A.S. Satyaningtijas Achmad Selamet Aku Adi Winarto Adisti Dwijayanti Adkhilni Utami Adkhilni Utami Adrian Situmeang Adrian Situmeang Adrian Situmeang Adrien Jems Akiles Unitly Agus Harsoyo Agus Harsoyo Agus Oman Sudrajat Agus Setiadi Al Azhar Al Mukhlas Fikri AL-AZHAR AL-AZHAR Alfred O. M. Dima Alif Iman Fitrianto Alif Iman Fitrianto Alkaustariyah Lubis Amrozi - Andri Maruli Tua Lubis Andri Maruli Tua Lubis Andriyanto A AUCKY HINTING Bambang Kiranadi Bayu Rosadi Berry Juliandi Bibiana W Lay BIBIANA W LAY Boenjamin Setiawan Boenjamin Setiawan Budiariati, Vista Budiono, Dwi Cece Sumantri Chairun Nisa Citra Noviana Cutnya’ Shaliran Nazlie (Alm) Dedy D. Solihin Diah Nugrahani Pristihadi Dian Anggraini Djaswadi Dasuki Djaswadi Dasuki Djoko Walujo Dody Dharmawan Trijuno Dondin Sajuthi Dwi Haryadi Elpita Tarigan EVY AYU ARIDA Evy Ayu Arida Farid A. Moeloek Ferry Sandra Frans Dhyanagiri Suyatna Frans Dhyanagiri Suyatna Frans Dhyanagiri Suyatna Hadi, Restu S. Handina Rakhmawati Handina Rakhmawati Harry Murti Harry Murti Harry Murti Harry Murti Hartanti Kusumaningtyas HERA MAHESHWARI HERDIS Herdis . herdis herdis Heri Sujoko Heru Setijanto I Gusti Agung Komang Diafari Djuni Hartawan I Ketut Mudite Adnyana I Ketut Suatha I Wayan Batan I Wayan Batan Ichsan Ichsan Iis Diatin Iman Supriatna Indra Bachtiar Indra Kusuma Irma H Suparto Irma Herawati Suparto Irma Suryani Ita Djuwita Ita Djuwita Ita Fauzia Hanoum Ita Fauzia Hanoum, Ita Fauzia Karisma Mardatillah KARTINI ERIANI Kartini Eriani KARTINI ERIANI Kartiwa, R. Angga Kelvin Yaprianto Kelvin Yaprianto Kelvin Yaprianto Ketut Adnyane Mudite Krido Brahmo Putro Kusdiantoro Mohamad Kusumaningtyas, Hartanti Latifah Kosim Darusman Lea Tarliyah M Agus Setiadi Madihah Madihah Madihah Madihah, Madihah Maman Surachman Min Rahminiwati Miraprahesti, Retti N. Mohamad Fakhrudin Mohammad Ghozali Mokhamad Fahrudin MOZES R. TOELIHERE Muhamad Haviz MUHAMMAD AGUS SUPRAYUDI Muhammad Gunawan Muhammad Gunawan Muhammad Haviz Muhammad Rizal Muhammad Rizal MUHAMMAD RIZAL MUHAMMAD RIZAL' Muhammad Rosyid Ridlo Muhammad Zairin JR Muhammad Zairin Jr. MULYOTO PANGESTU MULYOTO PANGESTU Mulyoto Pangestu Muslim Muslim Nastiti Kusumorini Nastiti Kusumorini Nazlie (Alm), Cutnya’ Shaliran Nining Handayani Nining Handhayani Noer Muhammad Dliyaul Haq Nurhidayat - Nurhidayat Nurhidayat Nuril Farizah Nursanti, Risa Nursanti, Risa R. Sapto Hendri Boedi Soesatyo Rachmat Herman Rahmaniyah, Wiwit Ridhani Ramadhan Sumarmin Rangga Setiawan Ratih Rinendyaputri Ratih Rinendyaputri Ratih Rinendyaputri Resti Rahma Dianti Ridi Arif Rimayanti - Rini Widyastuti Riris L. Puspitasari Riris L. Puspitasari Ronny Rachman Noor Sandy Qlintang SATRIYAS ILYAS Satya Gunawan Seiichi Watanabe Shofwal Widad Shofwal Widad Sigit Prastowo Siti Darodjah Rasad Sony H. Sumarsono SONY HERU SUMARSONO SONY HERU SUMARSONO Sri Catur Setyawatiningsih Sri Catur Setyawatiningsih Srihadi Agungpriyono Subangkit, Mawar Sulistiono Sumarsono, Sony H. Sumarsono, Sony Heru Supar - Supar . Sutarya Enus SUTIMAN BAMBANG SUMITRO Syamsunarno, Mas Rizky Anggun Adipurna TAKDIR SAILI Takdir Saili Takdir Saili TAKDIR SAILI TARUNI SRI PRAWAST MIEN KAOMINI ANY ARYANI DEDY DURYADI SOLIHIN Taufik Jamaan Thomas Mata Hine Thomas Mata Hine Trevino A. Pakasi TRINIL SUSILOWATI Tutik Wrediati Tutik Wresdiyati - Tutty Laswardi Yusuf Tuty Laswardi Yusuf Vincentia Maria Wahono Esthi Prasetyaningtyas Wahono Esti Prasetyaningtyas Wahono Esti PrasetyoningtyaserB Wahyudin Wasmen Manalu Widjiati w Wildan Mubarok Wining Astini Wining Astini Wiwit Ridhani Rahmaniyah Yoga Yuniadi Yuhara Sukra Yulnawati . YULNAWATI YULNAWATI Yundari, Yundari Yushinta Fujaya