Wahono Esthi Prasetyaningtyas
DepartemenAnatomi, Fisiologi dan Farmakologi, Fakultas Kedokteran Hewan, Institut Pertanian Bogor, Bogor

Published : 9 Documents
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Karakteristik dan komposisi semen kancil (Tragulus javanicus) yang dikoleksi dengan elektroejakulator

Jurnal Anatomi Indonesia Vol 1, No 1 (2006)
Publisher : Jurnal Anatomi Indonesia

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Abstract

The ejaculates were taken by electroejaculation from four apparently healthy young adultmale lessermousedeer (Tragulus javanicus). The animals were anesthetized with a combination of xylazin and ketamine followedby ether per inhalation anesthesia. The semen was white, yellowish, or creamy in color. The semen had a pH of7.63±0.22. Themean values for volumewas 19.44±6.8 μl, spermconcentration was 47.44 ± 4.9 x 106 sperm/ml,percentage of sperm motility was 36.43±1.1 %, percentage of live sperm was 53.11±3.0 %, percentage ofabnormal spermatozoa was 21.03±1.05%and percentage of intact acrosome was 52.28±2.7%, respectively.These values were relatively lowwhen compared to other domestic ruminants and suggested to be relatedwithage and sexualmaturity of the animal.The seminal plasma contained 10.2–11.5mg/100ml fructose, 22.07–24.5mg/100ml citric acid, 65mg/100ml proteins, 22.07–24.5mg/100ml sorbitol, 91.1– 94.72mg/100ml sodium, 0.1mg/100ml potassium, 12.8mg/100ml calcium, 0.8mg/100mlmagnesiumand 10.72–11.2 chloride. By SDS PAGE,eleven proteins with different molecular weight were determined in the seminal plasma. Among them, theproteins with 64.77 kDa and 71.72 kDa were particularly prominent.

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

Status of ram spermatozoa DNA after freeze-drying process

Jurnal Ilmu Ternak dan Veteriner Vol 11, No 3 (2006): SEPTEMBER 2006
Publisher : Indonesian Center for Animal Research and Development (ICARD)

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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

Status of ram spermatozoa DNA after freeze-drying process

Indonesian Journal of Animal and Veterinary Sciences Vol 11, No 3 (2006)
Publisher : Indonesian Animal Sciences Society

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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

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

Show Abstract | 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

Status of ram spermatozoa DNA after freeze-drying process

Jurnal Ilmu Ternak dan Veteriner Vol 11, No 3 (2006): SEPTEMBER 2006
Publisher : Indonesian Animal Sciences Society

Show Abstract | Original Source | Check in Google Scholar | Full PDF (260.586 KB)

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

ANNALES BOGORIENSES Vol 22, No 1 (2018): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

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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.

EVALUATION OF RAT LEYDIG CELL CULTURE COLLECTED WITH NYCODENZ GRADIENT IN PRODUCING TESTOSTERONE IN VITRO

Jurnal Kedokteran Hewan Vol 11, No 4 (2017): J. Ked. Hewan
Publisher : Syiah Kuala University

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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) (P<0.05). ITS and hCG in medium significantly increased Leydig cells concentration (10.40x106 cells/mL) at day 3 of culture (P<0.05). 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.

PF-17 The Development of Crude Testicular Cells in In Vitro Culture

Hemera Zoa Proceedings of the 20th FAVA & the 15th KIVNAS PDHI 2018
Publisher : Hemera Zoa

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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.