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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.
POLA DISTRIBUSI MITOKONDRIA SEL-SEL TROFOBLAS BLASTOSIS MENCIT (Mus Muculus Albinus) DAN PENGARUHNYA TERHADAP KEGAGALAN HATCHING DAN IMPLANTASI Helmita, Roza; Djuwita, Ita; Purwantara, Bambang
Sainstek : Jurnal Sains dan Teknologi Vol 7, No 1 (2015)
Publisher : IAIN Batusangkar

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (526.687 KB) | DOI: 10.31958/js.v7i1.121

Abstract

Implantation is the most critical stage in the establishment of pregnancy. In mammals, it has been estimated that between 25% and 60% of conceptuses are lost before or at the time of implantation. The objectives of  this study were to investigate  the ability of blastocyst hatching and attachment of trophoblast cells in in vitro, the outgrowth and differentiation of trophoblast cells in in vitro culture of hatched and non hatched blastocyst, the activity of mitochondria NADH-CoQ reductase and the pattern of mitochondrial distribution. Blastocysts were collected from mice cornua utery at day-4 of pregnancy and were divided into 3 groups: blastocysts undergo hatching within 24 hours, 48 hours and non hatching. Embryos were cultured in DMEM medium in 5% CO2 incubator at 37°C for 10 days. The trophoblasts monolayer were processed for Giemsa staining and histochemistry analysis of NADH-tetrazolium reductase activity. The outgrowth of trophoblast cells were measured using eyespiece micrometer. The results showed the ability of blastocyst hatching in in vitro were different. The trophoblast outgrowth diameter of 24 h hatched blastocyst was significantly different with the 48 h hatched and non hatched blastocyst. The activity of NADH-CoQ reductase of 24 h and 48 h hatched blastocysts showed higher intensity than the non hatched blastocysts (P<0,05) and the distribution of mitochondrial in trophoblast cell cytoplasma of 24 h and  48 h hatched blastocyst were homogen around nucleus whereas thoes of non hatched blastocyst were cluster and heterogen. In conclusion, in in vitro study the failure of blastocysts hatching and implantation was due to the failure of outgrowth and differentiation of the trofoblast cells and the impairment of NADH-CoQ reductase activity in complex I mitochondria to produce energy and the pattern of mitochondrial distribution.Key words: mitochondrial distribution, NADH- tetrazolium reductase, trophoblast, implantation 
PRIMORDIAL FOLLICLES DEVELOPMENT OF IMMATURE MICE OVARY AFTER FSH AND OVARY CUTTING TREATMENTS Djuwita, Ita
Jurnal Kedokteran YARSI Vol 18, No 1 (2010): JANUARI - APRIL 2010
Publisher : Lembaga Penelitian Universitas YARSI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33476/jky.v18i1.174

Abstract

The aims of the study were to investigate the influence of FSH and the ovary cutting treatments on the development of primordial follicles into preantral and antral follicles. Ovaries were grouped as whole ovary (without cutting); partially cut ovary and completely cut ovary (hemi ovary). All ovary groups were cultured in Dubellco?s Modified Eagle Medium (DMEM) containing 5ug/ml insulin, 10ug/ml transferrin, 5ug/ml selenium (ITS), 5% FBS, 50ug/ml gentamycin with and without 100uIU/ml Follicle Stimulating Hormone (FSH). Cultures were done in 5% CO2 incubator at 37oC. Results showed that after 8 days in vitro cultured in DMEM containing FSH, the average number of preantral follicles isolated from each whole, partially-cut and completely-cut ovaries were 16.1 + 3.3, 26.8 ± 7.7 and 12.3 + 1.9, respectively. On the other hand, those cultured in DMEM without FSH they were 17.3 + 3.8, 23.3 ± 5.2 and 17.8 + 2.8, respectively. After additional cultured for 8 days, the percentage of preantral follicles developing into the antral follicles in DMEM with and without FSH were 26.7 + 5.7 and 11.7 + 2.9, respectively. In conclusion, the supplementation of FSH in the culture medium did not increase the number of preantral follicles, but significantly increased the number of antral follicles. The ovary cutting treatment significantly increased the average number of collected preantral follicles.
AKTIVITAS ANTIFUNGAL MINYAK ATSIRI JINTEN PUTIH TERHADAP Candida parapsilosis SS25, C. orthopsilosis NN14, C. metapsilosis MP27, DAN C. etchellsii MP18 Ridawati, Ridawati; Jenie, Betty Sri Laksmi; Djuwita, Ita; Sjamsuridzal, Wellyzar
Makara Journal of Science Vol. 15, No. 1
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Antifungal Activity of Cumin Oil Against Candida parapsilosis SS25, C. orthopsilosis NN14, C. metapsilosis MP27, and C. etchellsii MP18. Many kinds of spices are used in Indonesia, one of them is white cumin seed. This spice is used not only for cooking, but also for traditional medicine. This study reported of antifungal activity from white cumin’s essential oil. Extraction and identification of Cumin oil were carried out. We obtained 2.5-3.0% of white essential oil which was colorless or light yellow color. GCMS analysis revealed that there were 12 peaks. Based on peak’s intensity the oil were dominated by 4 compound i.e. cuminaldehide (35.44%), ρ-cymene (34.77%), β-pynene (15.08 %) and γ-terpinene (8.15%). Growth inhibition zone determination has been carried out by diffusion disc and direct method against yeast i.e. C. parapsilosis SS25, C. orthopsilosis NN14, C. metapsilosis MP27, and C. etchellsii MP18. The results showed that all of the yeasts were sensitive to cumin oil. The inhibition zone radius were 13.4-16.5 mm. The cumin oil showed the inhibition of yeast growth with MIC values of 0.028%-0.042% and MFC values 0.09%- 0.14%, while nystatin had MIC values 0.40%-0.50% and MFC values 3.0%-4.0%. The activity of cumin oil was very strong as antifungal.
POTENSI TRANSDIFERENSIASI SEL FIBROBLAS MENJADI SEL SARAF SECARA IN VITRO (Transdifferentiation Potency of Fibroblast Cell to Neuron Cell in Vitro) Kaiin, Ekayanti M.; Djuwita, Ita
Jurnal Kedokteran Hewan Vol 10, No 1 (2016): March
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v10i1.3370

Abstract

The aim of this study was to examine the potency of fibroblast cells transdifferentiated to neuron cells in vitro. Newborn rat neuron conditioned medium (NBRN CM) was collected from neuron cells cultured with mDMEM without serum for 48 hours. Fibroblast cells were collected from fetal rat muscle treated with trypsin. Fibroblast cells were culture with 3 kind of culture medium: mDMEM + 0.01 mM -mercaptoethanol; mDMEM + 50% NBRN-CM and mDMEM + 0.01 mM -mercaptoethanol + 50% NBRN CM . As control, cells was cultured with mDMEM +10% newborn calf serum (NBCS). The addition of NBRN CM into culture medium resulted in 12.97% newborn cells in fibroblast culture medium passage I. Newborn rat neuron conditioned medium in fibroblast culture medium resulted 12.97% neuron cells at passage 1. The percentage was increased (14.60%) when - mercaptoethanol added into medium. The same result was found at passage 3 (12.67%; 13.17%). It showed that fibroblast cells has potency to transdifferentiated into neuron cells when cultured with NBRN CM. Further research is needed to know the fibroblast transdifferentiation potency.Key words: fibroblast, transdifferentiation, conditioned medium, neuron cells, in vitro
RESPONS SEL EPITEL USUS (CMT-93) TERHADAP NUTRASETIKAL GALOHGOR Roosita, Katrin; R, Rimbawan; Djuwita, Ita; Damanik, M. Rizal; Kusharto, Clara M.
Jurnal Kedokteran Hewan Vol 9, No 2 (2015): September
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v9i2.2825

Abstract

Penelitian ini bertujuan membandingkan pengaruh nutrasetikal galohgor dalam bentuk serbuk (GS) dan ekstrak (GE) terhadap proliferasi, morfologi, dan ekspresi gen Aldh1a2 pada sel epitel usus (CMT-93). Galohgor serbuk (GS) dibuat dengan menghancurkan semua bahan dandikeringkan menggunakan drum-dryer sedangkan GE dibuat dengan mengeringkan semua bahan dengan oven, digiling, dan dimaserasi dengan etanol selama 3x24 jam. Perlakuan didasarkan pada konsentrasi akhir -karoten yang berasal dari GS dan GE masing-masing sebesar 0,5; 1,5; dan 5,0 M dalam larutan medium Dulbecco's Modified Eagle's medium (DMEM) yang dilengkapi serum (10%). Analisis proliferasimenggunakan asai 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dan ekspresi gen dianalisis dengan reverse transcriptasepolymerase chain reaction (RT-PCR). Hasil penelitian meunjukkan bahwa GE pada konsentrasi tinggi (5,0 M) secara signifikan (P0,05) dapatmenekan proliferasi dan memengaruhi morfologi sel CMT-93. Beta-karoten dalam GE dan GS memengaruhi ekspresi gen Aldh1a2 pada sel epitel usus CMT-93
SEBARAN GLYCOCONJUGATE PADA SEL EPITEL OVIDUK KANCIL (Tragulus javanicus) H, Hamny; Agungpriyono, Srihadi; Djuwita, Ita; Wahyuni, Sri; Esthi Prasetyaningtyas, Wahono; Nasution, Idawati; Novelina, Savitri
Jurnal Kedokteran Hewan Vol 8, No 2 (2014): September
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | 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 m. Jenis lektin yang digunakan adalah biotinylated (Con A, PNA, RCA, UEA I, dan WGA) dengan dosis masing-masing sebanyak 15 g/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.
KONSENTRASI, KEMURNIAN, DAN VIABILITAS SEL LEYDIG HASIL PURIFIKASI DENGAN GRADIEN NYCODENZ DAN KULTUR IN VITRO M. Kaiin, Ekayanti; Djuwita, Ita; Yusuf, Tuty L.; Setiadi, M. Agus
Jurnal Kedokteran Hewan Vol 7, No 1 (2013): March
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v7i1.580

Abstract

Penelitian ini bertujuan isolasi dan purifikasi sel Leydig dengan menggunakan gradien Nycodenz untuk meningkatkan jumlah perolehan selLeydig yang hidup setelah purifikasi dan kultur. Isolasi dan purifikasi sel Leydig dilakukan dengan perlakuan gradien Nycodenz 3 kolom (5, 10, dan 15% sebagai Nycodenz I) dan 5 kolom (4, 8, 10, 12, dan 15% sebagai Nycodenz II) serta gradien Percoll 5 kolom (21, 26, 34, 40 dan 60%) sebagai kontrol. Parameter yang diamati adalah konsentrasi, kemurnian, dan viabilitas sel Leydig setelah isolasi dan purifikasi serta kultur in vitro. Hasil yang diperoleh menunjukkan bahwa isolasi dan purifikasi sel Leydig dengan gradien Nycodenz (I dan II) secara nyata (P0,01) menghasilkan konsentrasi sel yang lebih rendah dibandingkan dengan gradien Percoll. Namun penggunaan gradien Nycodenz II c enderungmenghasilkan kemurnian sel yang lebih tinggi (91,40%) dibandingkan dengan Nycodenz I (85,53%). Hasil tersebut tidak berbeda nyata dibandingkan dengan gradien Percoll (92,20%). Viabilitas sel Leydig pada semua perlakuan hampir sama yaitu 98%. Namun demikia n, setelah dikultur selama 3 hari, konsentrasi sel Leydig pada perlakuan Percoll (2,44x106 sel/ml) dan Nycodenz I (3,21x106 sel/ml) secara statistik (P0,05) lebih rendah dan dibandingkan dengan Nycodenz II (3,88x10 6 sel/ml), sedangkan viabilitas sel Leydig setelah dikultur pada gradien Nycodenz I (90,00%) dan Nycodenz II (91,17%) secara sangat nyata (P0,01) menghasilkan viabilitas yang lebih tinggi dibandingkan dengan gradien Percoll (82,30%). Berdasarkan hasil tersebut dapat disimpulkan bahwa penggunaan gradien Nycodenz II efektif untuk mempurifikasi sel Leydig dan setelah dikultur menghasilkan konsentrasi dan viabilitas sel yang lebih tinggi dibandingkan dengan gradien Percoll.
PRODUKSI EMBRIO KUCING SECARA IN VITRO DARI SPERMATOZOA HASIL PRESERVASI MELALUI FERTILISASI MIKRO Eriani, Kartini; Sukra, Yuhara; Boediono, Arief; Djuwita, Ita; Sumarsono, Sony Heru
Jurnal Kedokteran Hewan Vol 7, No 1 (2013): March
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v7i1.563

Abstract

Penelitian ini bertujuan mengetahui motilitas dan kemampuan memfertilisasi sperma kucing yang berasal dari epididimis yang disimpanpada suhu 4 C. Epididimis disimpan dalam media phosphate buffer saline (PBS) pada suhu 4 C selama 1, 3, dan 6 hari. Viabilitas spermatozoa diamati dengan pewarnaan hoechst-propidium iodine. Fungsi biologis spermatozoa dievaluasi melalui teknik kultur in vitro dengan fertilisasi mikro dan perkembangan embrio di dalam media kultur CR1aa. Hasil penelitian menunjukkan persentase spermatozoa hidup pada hari ke-1, 3, dan 6 penyimpanan masing-masing adalah 81,0; 71,7; dan 70,7% (duktus deferens), 84,0; 81,2; dan 63,2% (kauda epididimis), 84,0%; 75,0; dan 74,7% (korpus epididimis). Persentase pronukleus (PN) yang didapat dengan teknik intra cytoplasmic sperm injection (ICSI) menggunakan spermatozoa epididimis pada hari ke-1, 3, dan 6 hari penyimpanan masing-masing adalah 8,0; 10,0; dan 5,9%. Preservasi epididimis pada suhu 4 C dalam PBS dapat digunakan untuk fertilisasi dan menghasilkan embrio kucing secara in vitro.
PROLIFERASI DAN DIFERENSIASI SEL TULANG TIKUS DALAM MEDIUM KULTUR IN VITRO YANG MENGANDUNG EKSTRAK BATANG Cissus quadrangula Salisb. (SIPATAH-PATAH) Djuwita, Ita; Pratiwi, Irma Amalia; Winarto, Adi; Sabri, Mustafa
Jurnal Kedokteran Hewan Vol 6, No 2 (2012): September
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v6i2.295

Abstract

Penelitian mengenai pengaruh ekstrak batang Cissus quadrangula Salisb. (sipatah-patah) terhadap tingkat proliferasi dan diferensiasi sel-sel tulang tikus (Sprague Dawley) prepuber umur empat minggu menggunakan sistem kultur in vitro. Sel-sel tulang dikultur dalam medium dulbeccos modified eagles medium (DMEM) yang diberi tambahan newborn calf serum (NBCS) 10%, non essential amino acid (NEAA) 10%, NaHCO3, ITS 1 l/ml (mengandung insulin 5 g/ml, transferin 10 g/ml, selenium 5 g/ml; Sigma St Louis USA), dan 50 g/ml gentamisin (mDMEM), dengan dan tanpa ekstrak Cissus quadrangula (CQ). Penelitian terdiri atas lima perlakuan yakni kontrol positif (mDMEM + deksametason 10-8 M), kontrol negatif (mDMEM), dan tiga konsentrasi CQ: mDMEM + CQ 0,3 mg/ml; mDMEM + CQ 0,6 mg/ml; dan mDMEM + CQ 1,2 mg/ml. Kultur dilakukan dalam inkubator CO2 5%, pada suhu 37 C selama tujuh hari. Parameter yang diamati adalah konsentrasi sel, komposisi, diameter osteoblas, dan diameter osteosit. Konsentrasi sel dihitung menggunakan hemositometer Newbauer. Komposisi osteoblas dan osteosit ditentukan berdasarkan pengamatan morfologi di bawah mikroskop cahaya. Diameter sel diukur menggunakan eyepiece micrometer. Data dianalisis menggunakan analisis varians dan uji Duncan. Hasil penelitian menunjukkan bahwa pemberian ekstrak Cissus quadrangula Salisb. pada konsentrasi 0,3 mg/ml; 0,6 mg/ml; dan 1,2 mg/ml secara signifikan dapat meningkatkan proliferasi sel tulang; dan pada konsentrasi 0,6 mg/ml mampu menginduksi diferensiasi osteoblas menjadi osteosit (P0,05). Disimpulkan bahwa pemberian ekstrak Cissus quadrangula pada konsentrasi 0,6 mg/ml ke dalam medium kultur dapat meningkatkan proliferasi dan diferensiasi osteoblas.