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Status of ram spermatozoa DNA after freeze-drying process Saili, Takdir; Prasetyaningtyas, wahono Esthi; Setiadi, Mohamad Agus; AgungPriyono, Srihadi; Boediono, Arief
Indonesian Journal of Animal and Veterinary Sciences Vol 11, No 3 (2006)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (260.586 KB) | DOI: 10.14334/jitv.v11i3.528

Abstract

The process of freeze drying caused detrimental effect on plasma membrane and acrosome of the spermatozoa, even it potentially could alter the chromatin and DNA integrities. On the other hand, DNA integrity is essential for spermatozoa to participate in pronucleus formation during fertilization event. Therefore the evaluation of DNA integrity should be carried out to study the effect of freeze drying process. EDTA, EGTA, and PBS were used as dilution media of spermatozoa prior to freeze drying process to protect the DNA. Toluidine blue staining and comet assay methods were used to evaluate the alteration on chromatin and DNA integrities of spermatozoa, respectively. The results revealed that the highest compacted chromatin after 6 months storage of freeze-dried spermatozoa were observed from EGTA-3 (98%) and EGTA-1 (97%) treatments that had significant differences compared to all PBS treatments (90-92%), but not for fresh spermatozoa (100%). Whereas, the highest compacted DNA integrity of freeze-dried spermatozoa were observed from EGTA-2 (92%) and EGTA-3 (92%) but had no significant differences compared to other treatments including fresh spermatozoa (97%). These results demonstrate that EDTA and EGTA tend to be able to protect chromatin and DNA integrities of ram spermatozoa during freeze-drying and storage compared to PBS. Key Words: Freeze-Drying, Spermatozoa, DNA, Toluidine Blue, Comet Assay
Biological Analysis of Leydig Cells-Conditioned Medium To Support Rat Bone Marrow Mesenchymal Stem Cells Differentiation Kaiin, Ekayanti Mulyawati; Prasetyaningtyas, Wahono Esthi; Mohamad, Kusdiantoro; Djuwita, Ita; Yusuf, Tuty Laswardi; Setiadi, Mohamad Agus
ANNALES BOGORIENSES Vol 22, No 1 (2018): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ab.v22i1.328

Abstract

The developed Leydig cells-conditioned medium (LCM) contains bioactive materials secreted by Leydig cells in vitro.  LCM was used to evaluate the ability of bone marrow mesenchymal stem cells differentiation. Bone marrow mesenchymal stem cells (1x 106 cell/ml) were cultured in : 1) DMEM supplemented with 10% NBCS as a control (M), 2) M supplemented with 10 ng/ml testosterone; 3) M supplemented with 50%  LCM ; 4) M supplemented with 50% LCM and 2.5 IU/ml hCG. Bone marrow mesenchymal stem cells that were cultured with  LCM has a positive reaction (57.4%) to histochemistry staining 3β-HSD and produced 1.87 ng/ml testosterone. Supplementation of hCG to LCM  increased the positive number of Leydig cells and testosterone production by 74.6% and 12.33 ng/ml (P<0.05). It can be concluded that Leydig cells-conditioned medium can support differentiation of bone marrow mesenchymal stem cells into Leydig cells.
Allotransplantasi Testis Mencit Muda sebagai Upaya Preservasi Gonad In Vivo Wahono Esthi Prasetyaningtyas; Kusdiantoro Mohamad; Mokhamad Fahrudin; Ita Djuwita; Srihadi Agungpriyono
Jurnal Ilmu Pertanian Indonesia Vol. 12 No. 1 (2007): Jurnal Ilmu Pertanian Indonesia
Publisher : Institut Pertanian Bogor

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (166.442 KB)

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
PF-17 The Development of Crude Testicular Cells in In Vitro Culture Wahono Esthi Prasetyaningtyas; Ni Wayan Kurniani Karja; Srihadi Agungpriyono; Mokhamad Fahrudin
Hemera Zoa Proceedings of the 20th FAVA & the 15th KIVNAS PDHI 2018
Publisher : Hemera Zoa

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (488.568 KB)

Abstract

Spermatogenesis is a continuous process in which spermatogonial stem cells (SSC) develop into specific germ cells before terminally differentiating to form spermatozoa.  The process is supported by Sertoli cells, which are in close contact with germ cells in the seminiferous tubules. Sertoli cells provide essential hormonal signals, nutrients, and physical support to germ cells for successful spermatogenesis.The crude testicular cells (CTC) contains many cell types, like Sertoli cell, Leydig cell, spermatogonial stem cell (SSC), spermatocyte and other testicular somatic cells (Shah et all. 2016). Testicular cells are believed to secrete various growth factors that induced the spermatogenesis process.  The spermatogonial stem cells are unique population of cells in the male testis, which dual function.  First self-renewing their population to maintain the number of stem cells, secondary function is differentiating into spermatids in testis (Wang et al.  2015).Spermatogenic cells differentiation  needed the similar microenvironment in vivo spermatogenesis.  The essential nutrients was collected from healty culture and the culture contained mixed population of cells both the somatic cells and spermatogenic cells.  To identification the spermatogenic cells using Periodic Acid Schifft (PAS) staining (Chang et al. 2011). The present study examined the development of crude testicular cells using PAS staining.
AH-25 Sperm Morphology of the Javan Muntjak, Muntiacus muntjak muntjak Sri Wahyuni; Gholib Gholib; Wahono Esthi Prasetyaningtyas; I Ketut Mudite Adnyane; Srihadi Agungpriyono; Hamny Hamny; Muhammad Jalaluddin; Mustafa Sabri; Muslim Akmal; Muhammad Agil; Tuty Laswardi Yusuf
Media Veteriner Proceedings of The 5th Congress of Asian Association of Veterinary Anatomists (Asian AVA) 2015
Publisher : Media Veteriner

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Abstract

Sperm Morphology of the Javan Muntjak,  Muntiacus muntjak muntjak
Efektivitas Low Density Lipoprotein (LDL) dari Kuning Telur Ayam terhadap Kualitas Semen Cair Domba Dwitya Citraesti; Wahono Esthi Prasetyaningtyas; Ni Wayan Kurniani Karja
Jurnal Sain Veteriner Vol 39, No 3 (2021): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.63395

Abstract

Low-Density Lipoprotein (LDL) extracted from egg yolk (LDL) has recently known can eliminate the adverse effect associated in the use of fresh egg yolk. The role of LDL in liquid preservation at 4°C of ram sperm has not been explored. This research evaluates the effects of substituting egg yolk with LDL in ram sperm preservation at 4 °C on 5 days. The objective of this research was to assess the effects of substituting egg yolk with LDL for use as an extender in sperm preservation at 4°C, as well as on spermatozoa motility, viability, morphology, plasma membrane, and acrosome integrity, for 5 days. The semen was subsequently divided into five and diluted with Tris–fresh egg yolk (K), Tris–LDL5% (LDL5), Tris–LDL10% (LDL10), Tris–LDL15% (LDL15), and Tris–LDL20% (LDL20). The result showed a significant difference between LDL to fresh egg yolk for ram sperm quality (P<0.05). The effectiveness of LDL on sperm quality decreased following by its concentration. Even though up to 20% concentration of LDL, it can not preserve the quality of diluted semen for motility, viability, and plasm membrane integrity. 
EVALUATION OF RAT LEYDIG CELL CULTURE COLLECTED WITH NYCODENZ GRADIENT IN PRODUCING TESTOSTERONE IN VITRO Ekayanti Mulyawati Kaiin; Wahono Esthi Prasetyaningtyas
Jurnal Kedokteran Hewan Vol 11, No 4 (2017): December
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (285.769 KB) | DOI: 10.21157/j.ked.hewan.v11i4.3984

Abstract

 The aim of this study was to evaluate the ability of rat’s Leydig cells collected with Nycodenz gradient in producing testosterone in vitro. Leydig cells were collected using 5 column of Nycodenz gradient (4, 8, 10, 12, and 15%) and cells were evaluated regarding its concentration, viability, and purity of Leydig cells. Media used to cultured Leydig cells were Dulbecco’s Modified Eagle’s Medium (DMEM)+10% newborn calf serum (NBCS); DMEM+10% NBCS+2,5 IU/mL human chorionic gonadotrophin (hCG); DMEM+10% NBCS+ 5 µg/mL insulin, 10 µg/mL transferrin, and 5 µg/mL Se (ITS); DMEM+10% NBCS+hCG+ITS at 5% CO2 incubator with temperature of 37.5° C for 3 days. Culture medium was collected every day for testosterone analysis with enzyme-linked immunosorbent assay (ELISA). By adding ITS to the medium, Leydig cells concentration was significantly increased (8.92x106  cells/mL) compared to medium with serum (7.74x106 cells/mL) or hCG (7.68x106 cells/mL) (P0.05). ITS and hCG in medium significantly increased Leydig cells concentration (10.40x106 cells/mL) at day 3 of culture (P0.05). The result of parallelism test showed that the assay obtained good validity to measure testosterone concentration in culture medium. Testosterone in medium was detected at 1.80-2.60 ng/mL at day 1 of culture. In conclusion, Leydig cells collected with Nycodenz gradient had no effect to testosterone secretion from Leydig cells in vitro.
SEBARAN GLYCOCONJUGATE PADA SEL EPITEL OVIDUK KANCIL (Tragulus javanicus) Hamny H; Srihadi Agungpriyono; Ita Djuwita; Sri Wahyuni; Wahono Esthi Prasetyaningtyas; Idawati Nasution; Savitri Novelina
Jurnal Kedokteran Hewan Vol 8, No 2 (2014): September
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (290.597 KB) | DOI: 10.21157/j.ked.hewan.v8i2.2623

Abstract

Penelitian ini bertujuan mengetahui distribusi glycoconjugate yang terekspresi pada sel epitelium oviduk kancil (Tragulus javanicus). Dalam penelitian ini digunakan satu oviduk kancil yang berasal dari satu ekor kancil b etina dewasa berumur lebih dari satu tahun. Sampel difiksasi dengan larutan Bouin dan diproses menurut standar histologi sampai menjadi blok parafin dan dipotong dengan ketebalan 5 µm. Jenis lektin yang digunakan adalah biotinylated (Con A, PNA, RCA, UEA I, dan WGA) dengan dosis masing-masing sebanyak 15 µg/ml. Hasil penelitian diketahui bahwa glycoconjugate dengan residu gula galaktosa, glukosa, manosa, N-asetilgalaktosamin, N-asetilglukosamin, fukosa, dan asam sialat ditemukan pada bagian apikal sel epitel dan di dalam sitoplasma. Glycoconjugate dengan residu gula N-asetilgalaktosamin merupakan glycoconjugate yang paling banyak ditemukan di bagian apikal sel epitel dan di dalam sitoplasma dibandingkan dengan glycoconjugate dengan residu gula lainnya.
The Quality of Indo-Pacific Bottlenose Dolphin (Tursiops aduncus) Sperm Following Liquid-storage in Low Temperature Fahrudin, Mokhamad; Gusdinar, Rizal; Arifiantini, Raden Iis; Prasetyaningtyas, Wahono Esthi; Adnyane, I Ketut Mudite; Elmanaviean, Muhammad; Nugraha, Arifin Budiman; Karja, Ni Wayan Kurniani
ILMU KELAUTAN: Indonesian Journal of Marine Sciences Vol 29, No 4 (2024): Ilmu Kelautan
Publisher : Marine Science Department Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/ik.ijms.29.4.456-466

Abstract

Indo-Pacific bottlenose dolphin (Tursiops aduncus) is a marine mammal that lives in relatively small populations. The geographic ranges of this species are susceptible to the effects of human activities, thereby necessitating conservation efforts to prevent extinction. Therefore, this study aimed to evaluate the daily quality of dolphin sperm after several days of refrigeration. The sperm of two male dolphins were stored at 4oC for 4 days, and the quality was observed daily to determine the motility, viability, membrane integrity, and sperm abnormalities. Sperm samples were divided into four groups, consisting of two centrifuged followed by the removal of seminal plasma, and two groups without centrifugation, containing 100x106 and 200x106 sperm/ml each. After liquid storage, the motility of sperm was 63-75% with no significant reduction in the first 3 days. Sperm viability following storage was 65-75% and the percentage with abnormal morphology ranged from 2-6%. Furthermore, there was no significant increase in abnormal morphology of sperm on any day of storage for 3 days. Sperm membrane integrity was 36-49%, with no significant reduction in the membrane integrity in the first 2 days. There was no significant difference in sperm quality, although centrifugation and removal of seminal plasm had a slight effect. The results of this study showed that Indo-Pacific bottlenose dolphin sperm could be stored for a short period as liquid storage while maintaining a quality that allows for future use.
FROZEN SEMEN CHARACTERISTICS OF BALI BULL IN DIFFERENT AGE GROUPS Witri, Brilla Widya; Arifiantini, Iis; Prasetyaningtyas, Wahono Esthi; Karja, Ni Wayan Kurniani
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 12 No. 1 (2025)
Publisher : BRIN - Badan Riset dan Inovasi Nasional

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.55981/jbbi.2025.10087

Abstract

One factor that affects frozen semen characteristics is bull age. Increasing age induces changes in sperm that reduce frozen semen quality. This study aimed to evaluate the frozen semen characteristics of Bali bulls aged 3, 8, and 13 years. Frozen semen was derived from fresh semen with sperm motility > 70%. The frozen semen characteristics tested included motility, viability, plasma membrane integrity, acrosome integrity, and sperm abnormalities. Sperm motility was analyzed by computer-assisted sperm analysis (CASA). Viability was assessed using eosin-nigrosin staining; plasma membrane integrity was assessed using the hypoosmotic swelling test (HOST); acrosome integrity was assessed using Giemsa staining; and sperm abnormalities were assessed using Williams staining. The plasma membrane integrity of the 13-year-old group was significantly lower (P < 0.05) than that of the younger age groups. However, there were no significant differences (P > 0.05) in the other parameters. We conclude that age affects the integrity of the sperm plasma membrane in frozen semen from Bali bulls.