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

 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

Efektivitas Low Density Lipoprotein (LDL) dari Kuning Telur Ayam terhadap Kualitas Semen Cair Domba

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.

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

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

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

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

Morphological and lectin histochemical studies of the spermatozoa of the lesser mouse deer (Tragulus javanicus)

The morphology and lectin histochemical property of the spermatozoa of the lesser mouse deer (Tragulus javanicus) were studied using light and scanning electron microscopy. Spermatozoa were small in size and measured 36.52 ± 5.6 μm in length. The head of the spermatozoa was relatively round in shape (5.55 ± 0.8 μm in length and 4.77± 0.5 μm in width). Lectin peanut (Arachis hypogea) agglutinin (PNA) and lectin wheat germ (Triticum vulgaris) nagglutinin (WGA) were positive in the membrane of the acrosomal region, and in the membrane at the anterior part of the acrosomal region and in the tail, respectively. It is suggested that carbohydrates with galactose β1-3, D-N acetylgalactosamine, D-N acetylglucosamine and sialic acid sugar residues may be involved and may have significant roles in the spermatozoa in the lesser mouse deer

SEBARAN GLYCOCONJUGATE PADA SEL EPITEL OVIDUK KANCIL (Tragulus javanicus)

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

AH-25 Sperm Morphology of the Javan Muntjak, Muntiacus muntjak muntjak

Sperm Morphology of the Javan Muntjak,  Muntiacus muntjak muntja

MtDNA variation and human-mediated introgression of indigenous sus populations on several Indonesian Islands

o examine the genetic origin of the domestic pig, the distribution of wild boar, and　human-mediated translocation of the domestic pig, we collected 223 samples from domestic pigs and　wild boars from eight Indonesian islands, sequenced the control region of mitochondrial DNA　(mtDNA) from each sample, and compared these sequences with previously determined sequences from East and Southeast Asian domestic pigs and wild boars. Three Sus species (S. scrofa, S.verrucosus, and S. celebensis) were identified on the Indonesian islands. The mtDNA sequences of　three Indonesian Sus species were diverse, and they clustered into three lineages with low bootstrap　values (an S. scrofa group including East and Southeast Asian domestic pigs and wild boars, a group including indigenous S. scrofa together with S. verrucosus from Sumatra and Java Islands, and an S.celebensis group from Sulawesi Island). The mtDNA haplotypes of S. scrofa wild boars from three　(Sumbawa, Flores and New Guinea) islands and domestic pigs from two (Lombok and Timor) islands east of the Wallace Line, and some S. scrofa wild boars from Sumatra and Java Islands were related　to Vietnamese pig mtDNA sequences in the East and Southeast Asian domestic pig and wild boar　clade, supporting that ancient immigrants likely introduced domestic pigs from the Asian continent to east Indonesian islands. The mtDNA haplotypes of S. celebensis were broadly divided into three　groups, which were distributed in the north and southwest areas, central area and southeast area of　Sulawesi Island

ALLOTRANSPLANTASI TESTIS MENCIT MUDA SEBAGAI UPAYA PRESERVASI GONAD IN VIVO

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 patient

CARBOHYDRATES OF CHANGES DURING THE FOLLICULAR DEVELOPMENT IN THE OVARY OF THE MOUSE DEER, TRAGULUS JAVANICUS

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.</em