29 Aug 2014
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 Dulbeccoâs 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 sulfateâpolyacrilamide 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.
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