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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.
Analisis Proteomik Cairan Sinovial Sendi Domba: Efektivitas Metode dan Profil Protein Fungsional Mohamad, Kusdiantoro; Rahmaniyah, Wiwit Ridhani; Adnyane, I Ketut Mudite; Fahrudin, Mokhamad; Boediono, Arief
Acta VETERINARIA Indonesiana Vol. 8 No. 2 (2020): Juli 2020
Publisher : IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1046.317 KB) | DOI: 10.29244/avi.8.2.52-64

Abstract

Penelitian ini bertujuan untuk mengevaluasi metode deplesi dan digesi protein, serta menganalisis proteom cairan sinovial (SF) sendi pada domba sehat menggunakan kromatografi cair–tandem spektrometri massa. Cairan SF dikoleksi dari enam ekor domba garut betina, umur ±4 tahun, berat 35-40 kg, dan sehat secara klinis. Deplesi protein berkelimpahan tinggi dilakukan dengan metode spin column menggunakan TOP 12 dan metode proteospin, sementara digesi protein dengan tiga metode, yaitu digesi in solution, in gel, dan in-solution + filter-aided sample preparation (FASP). Metode yang terbaik kemudian dilanjutkan dengan fraksinasi peptida menggunakan ultra performance liquid chromatography. Metode deplesi proteospin dengan digesi protein in-solution + FASP merupakan metode terbaik berdasarkan nilai coverage dan sequest HT. Hasil analisis proteomik terkarakterisasi 52 protein pada database spesies domba. Anotasi gen ontologi menggunakan DAVID analysis menunjukkan bahwa protein-protein SF tersebut merupakan komponen sel terutama sebagai eksosom ekstraseluler, fungsi molekuler sebagai aktivitas inhibitor endopeptidase tipe-serin dan pengikat ion kalsium; serta proses biologis sebagai angiogenesis dan koagulasi darah atau pembentukan fibrin. Analisis KEGG pathway menunjukkan protein SF berperan utama pada jalur kaskade koagulasi dan komplemen.
Heterotopic grafting sites can be useful in producing oocytes for in vitro Fertilization, therefore, maximising the oocyte yield from the graft by gonadotrophin stimulation would be advantageous. The aim of this study was to investigate the number and quality of oocytes collected from heterotopic autografted ovary after Pregnant Mare Serum Gonadothropin (PMSG) induction. Graft recipients were treated either with or without PMSG stimulation 48 hours prior to graft collection. Ovarian tissue from . NURBARIAH; ITA DJUWITA; KUSDIANTORO MOHAMAD; IMAN SUPRIATNA
HAYATI Journal of Biosciences Vol. 18 No. 4 (2011): December 2011
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.18.4.151

Abstract

Heterotopic grafting sites can be useful in producing oocytes for in vitro Fertilization, therefore, maximising the oocyte yield from the graft by gonadotrophin stimulation would be advantageous. The aim of this study was to investigate the number and quality of oocytes collected from heterotopic autografted ovary after Pregnant Mare Serum Gonadothropin (PMSG) induction. Graft recipients were treated either with or without PMSG stimulation 48 hours prior to graft collection. Ovarian tissue from four weeks old mice (DDY strain) were autotransplanted under the kidney capsule of the same ovariectomized mice and the oocytes were collected 21 days after autotransplantation. The results showed that the average number of oocytes collected from autografted ovaries without PMSG induction were 9.0. + 2.8 not significantly different with those received PMSG induction, 10.9 + 5.1. The percentage of matured and fertilized oocytes and the developed embryos from the autografted ovaries without PMSG induction were 52.4, 33.4, and 26.0%, respectively not significantly different with those received PMSG induction, 53.2, 35.1, and 29.9%, respectively. The number of oocytes and the capacity to matured, fertilized and developed were significantly lower (P < 0.05) compared to the superovulated nongrafted (control) ovaries. In conclusion, PMSG induction on the graft recipients did not significantly increase oocytes yield from grafted heterotopic ovaries. The number and quality of oocytes produced from the autografted ovaries were lower than the superovulated nongrafted ovaries, but still can be used for in vitro embryo production after sequential in vitro maturation and fertilization.
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
Pemaparan Gelombang Elektromagnetik Telepon Genggam pada Mencit (Mus musculus albinus) Periode Awal Kebuntingan Vincentia Maria; Kusdiantoro Mohamad; Arief Boediono
Acta VETERINARIA Indonesiana Vol. 2 No. 1 (2014): Januari 2014
Publisher : IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (265.16 KB) | DOI: 10.29244/avi.2.1.36-41

Abstract

Peningkatan penggunaan telepon genggam diiringi dengan peningkatan kepedulian masyarakatterhadap bidang kesehatan dan keamanan terkait emisi gelombang elektromagnetik yang berasaldari telepon genggam. Penelitian ini bertujuan mengetahui tingkat keamanan paparan gelombangelektromagnetik telepon genggam melalui pengamatan terhadap jumlah implantasi dan anak lahirdengan menggunakan mencit sebagai hewan model. Dua puluh empat ekor mencit betina disinkronisasidan dikawinkan dengan mencit jantan dengan perbandingan jantan dan betina 1:1 (single mating).Pemaparan dilakukan dengan gelombang berfrekuensi 900 MHz selama 7 hari pertama setelah mencitkawin. Mencit betina dibagi ke dalam empat kelompok berdasarkan lamanya waktu paparan. Waktupaparan adalah satu, dua, dan empat kali per hari dengan masing-masing lama paparan 15 menit untukkelompok pertama, kedua, dan ketiga, sementara kelompok keempat sebagai kelompok kontrol tidakdiberikan paparan. Hasil penelitian menunjukkan tidak adanya pengaruh nyata lama paparan gelombangelektromagnetik telepon genggam terhadap jumlah implantasi dan anak lahir dari induk yang terpapar(P>0,05). Untuk seluruh kelompok, jumlah implantasi berkisar antara 8,66 sampai dengan 10,00 danjumlah anak lahir berkisar antara 10,00 sampai dengan 12,33; tidak berbeda nyata dengan nilai padakelompok kontrol. Berdasarkan hasil penelitian, lama paparan gelombang elektromagnetik yang berasaldari telepon genggam dalam penelitian ini masih berada dalam tingkat aman untuk mencit bunting.
Analisis Proteomik Cairan Sinovial Sendi Domba: Efektivitas Metode dan Profil Protein Fungsional Kusdiantoro Mohamad; Wiwit Ridhani Rahmaniyah; I Ketut Mudite Adnyane; Mokhamad Fahrudin; Arief Boediono
Acta VETERINARIA Indonesiana Vol. 8 No. 2 (2020): Juli 2020
Publisher : IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1046.317 KB) | DOI: 10.29244/avi.8.2.52-64

Abstract

Penelitian ini bertujuan untuk mengevaluasi metode deplesi dan digesi protein, serta menganalisis proteom cairan sinovial (SF) sendi pada domba sehat menggunakan kromatografi cair–tandem spektrometri massa. Cairan SF dikoleksi dari enam ekor domba garut betina, umur ±4 tahun, berat 35-40 kg, dan sehat secara klinis. Deplesi protein berkelimpahan tinggi dilakukan dengan metode spin column menggunakan TOP 12 dan metode proteospin, sementara digesi protein dengan tiga metode, yaitu digesi in solution, in gel, dan in-solution + filter-aided sample preparation (FASP). Metode yang terbaik kemudian dilanjutkan dengan fraksinasi peptida menggunakan ultra performance liquid chromatography. Metode deplesi proteospin dengan digesi protein in-solution + FASP merupakan metode terbaik berdasarkan nilai coverage dan sequest HT. Hasil analisis proteomik terkarakterisasi 52 protein pada database spesies domba. Anotasi gen ontologi menggunakan DAVID analysis menunjukkan bahwa protein-protein SF tersebut merupakan komponen sel terutama sebagai eksosom ekstraseluler, fungsi molekuler sebagai aktivitas inhibitor endopeptidase tipe-serin dan pengikat ion kalsium; serta proses biologis sebagai angiogenesis dan koagulasi darah atau pembentukan fibrin. Analisis KEGG pathway menunjukkan protein SF berperan utama pada jalur kaskade koagulasi dan komplemen.
The Influence of Homologous and Heterologous Sera in Culture Medium on in Vitro Maturation and Fertilization of Sheep Oocytes Ita Djuwita; Yohan Rusyiyantono; Kusdiantoro Muhamad; Bambang Purwantara; Yuhara Sukra
Media Veteriner Vol. 5 No. 1 (1998): Media Veteriner
Publisher : Media Veteriner

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (749.792 KB)

Abstract

The influence of various types of sera i.e. Fetal Calf Serum (FCS), Bovine Post Estrous Serum (SPES) as heterolog sera; and Ewes Fetal Serum (EFS) and Ewes Serum (ES) as homologue sera on in vitro maturation and fertilization of sheep oocytes was investigated using Tissue Culture Medium 199TM (TCM 199. Gibco). Sovine sera were collected from cow at seven days post-estrus, while ewes sera were collected during estrous (ES-HO) and six days post estrous (ES-H6). The result shows that the maturation rate of oocytes cultured in medium supplemented with EFS, ES-HO and ES-H6 at  concentrations of 32.9%, 68.7%, and 67.6% respectively is better than those supplemented with FCS and SPES at concentrations of 22.2% and 28.9%, respectively. In vitro fertilization rate were significantly higher in medium supplemented with ES-HO and ES-H6 at concentration of 65.4% and 65.8% (P < 0.05) than that supplemented with SPES at concentration of 31.7%. In conclusion, ewes sera could be used in culture medium for promoting both in vitro maturation and fertilization rate of sheep oocytes.
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

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (175.23 KB)

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

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (114.452 KB)

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

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (182.817 KB)

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.