Kultur In Vitro Sel-sel Fibroblas Fetus Tikus
Abstract
Research has been conducted on the in vitro culture of rat fetal fibroblast using Dulbeccos’ Modified Eagle Medium supplemented with 3mM sodium bicarbonate, 10% fetal calf serum and 50 μg/ml gentamycin (mDMEM) in CO2 incubator at 37oC. Evaluation was done on the proliferation rate and the protein production after several passages. The growth of the fibroblast was confirmed by their morphology. The proliferation rate was done based on the population doubling time (PDT) by counting the number of cells in each passage by using Improved Neubauer hemocytometer. Identification of proteins was done quatitatively using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). Based on morphological observation the cell grown in in vitro culture were the fibrocyte and fibroblast and showed increasing of homogenicity after several passages. Based on the PDT, the proliferation rate was increased by each passages. Fibroblast also secrete proteins remain until passage 9 based on the results of electrophoresis. It can be concluded that the growth and protein secretion of rat fetal fibroblast in in vitro culture can be maintained up to 9 passages.