ultur In Vitro Sel-sel Otak Besar (Cerebrum) Anak Tikus
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Research has been conducted on in vitro culture of three days old rat cerebrum cells in Dulbecco’s Modified Eagle Medium (DMEM) containing 10% Fetal Bovine Serum (FBS) and 50 μg/ml gentamycin, and supplemented with and without 5 μg/ml insulin, 10 μg/ml transferin, 5 μg/ml selenium (ITS). Culture was done in 5% CO2 incubator at 37oC for 11 days. Identification was done on types of cell based on morphological observation, growth capacity (cell proliferation, population doubling time/ PDT and length of axon and dendrit) based on calculation of the number of cells using hemocytometer and micrometer, and the protein produced by the cerebrum cells in the culture medium using sodium dodecyl sulfate–polyacrilamide gel electrophoresis (SDS-PAGE) method. Quantitative data were analyzed using statistical T-test on Minitab program. The results showed that based on the morphological observations, there are two types of cells including nerve cells of bipolar and multipolar neurons and glial cells including the fibrous astrocyte, protoplasmic astrocyte, oligodendrocyte, and microglia. Supplementation of ITS into the culture medium could increase the cells proliferation rate (P<0.05) with lower PDT, axon and dendrit lenght growth and quantitatively the 21.5–36.5 kDa protein production as indicated by the intensity of the protein band.