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Ita Djuwita
Bagian Klinik Hewan, Fakultas Kedokteran Hewan, Universitas Udayana, Bali
Veterinary

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Induksi Ekstrak Pegagan Secara in vitro terhadap Proliferasi dan Diferensiasi Sel-Sel Otak Besar Anak Tikus (IN VITRO INDUCTION OF CENTELLA ASIATICA (PEGAGAN) EXTRACT ON THE PROLIFERATION AND DIFFERENTIATION OF NEWBORN RAT CORTEX CEREBRI CELLS) Ita Djuwita; Min Rahminiwati; Latifah Kosim Darusman; Siti Sa’diah
Jurnal Veteriner Vol 14 No 2 (2013)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

The aim of the study was to analyze the potency of Centella asiatica extract to induce proliferation andneurogenesis process of newborn rats cortex cerebri.   Research has been conducted on in vitro culture ofthree days old rat (Sprague Dawley) cerebrum cells in DMEM (Dulbecco’s Modiûed Eagle’s Medium)containing 10% NEAA (Non Essential Amino Acid), 1 mM NaHCO3, 10% NBCS (Newborn Calf Serum)and 50 µg/mL gentamycin (mDMEM), with and without Centella asiatica (CA) leaf extracts. The experimentwas set in five groups of treatment consisted of positive control (mDMEM+30 µg/mL asiaticoside (AC)),negative control (mDMEM), and mDMEM with three concentration of CA extract i.e. 100 ppm, 200 ppmand 400 ppm. Culture was done in 5% CO2 incubator at 37oC for six days. The parameters observed werecells proliferation based on Population Doubling Time (PDT), neuron and glia composition, and the lengthof axon and dendrite. Cells concentration were counted using Newbauer hemocytometer.  Neuron and gliacells were determined based on morphology after Hematoxylin-Eosin staining, and the length of axon anddendrite were measured using eyepiece micrometer. Data were analyzed using ANOVA and Duncan test.The results showed that Centella asiatica extract at concentration 100 ppm could induce neurogenesisand increased the axon length growth.  However, at concentration 200 and 400 ppm, CA extract  inhibitedthe neuronal cells proliferation and the axonal growth (P<0,05). In conclusion, induction of Centella asiaticaextracts at concentration of 100 ppm on the cortex cell cerebrum cells culture increase the axon lengthgrowth and tends to induce neurogenesis; however at higher concentration CA extract was neurotoxic.
Pertumbuhan dan Sekresi Protein Hasil Kultur Primer Sel-Sel Serebrum Anak Tikus (IN VITRO GROWTH AND PROTEIN SECRETION OF NEWBORN RAT CEREBRAL PRIMARY CELLS CULTURE) Ita Djuwita; Vivit Riyacumala; Kusdiantoro Mohamad; Wahono Esthi Prasetyaningtijas; Nurhidayat .
Jurnal Veteriner Vol 13 No 2 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

The objective of this study was to identify the proliferation and differentiation of mice cerebrum cellsand its protein product. Research has been conducted on in vitro growth of three days old rat cerebrum cellsin Dulbecco’s Modified Eagle Medium (DMEM) containing 10% Amino Acid Non Essential (AANE), 10%Fetal Bovine Serum (FBS), 3mM NaHCO3, 50 mg/ml gentamycin, and supplemented with and without 5mg/ml insulin, 10 mg/ml transferin, 5 mg/ml selenium (ITS). Culture was done in 5% CO2, then incubatedat 37oC until 90% confluent. Identification were done on cell proliferation and population doubling time(PDT), number of neuron and glial cells, length of axon and dendrit, and protein secretion. The number andlength of neuronal cells were calculated by using hemocytometer and eyepiece micrometer, respectivelly.Protein secreted into culture medium, designed as conditioned medium (CM) was analysized using sodiumdodecyl sulfate–polyacrilamide gel electrophoresis (SDS-PAGE) method. Quantitative data were analyzedusing statistical T-test on Minitab program. In vitro culture of rat cerebrum cells showed two types of cellsincluding nerve cells of bipolar and multipolar neurons and glial cells including astrocyte (the fibrous andprotoplasmic), oligodendrocyte, and microglia. Supplementation of ITS into the culture medium increasedthe cell proliferation rate (P<0.05) with lower PDT, and quantitatively, the 30 kDa secreted protein asindicated by the higher intensity of the protein band.
Co-Authors Kusdiantoro Mohamad Latifah Kosim Darusman Nurhidayat . Rahminiwati, Min Siti Sa’diah Vivit Riyacumala Wahono Esthi Prasetyaningtijas