Paper ID: 13
IN VITRO FIBROBLAST CELLS CULTURE FROM PELUNG CHICKEN EMBRYO AND ITS POTENTIAL APPLICATION
Elisa Herawati1, *, Harlina Wulandari1, Shanti Listyawati1, Nita Etikawati1
Study Program of Biology, Faculty of Mathematics and Natural Science, Universitas Sebelas Maret, Surakarta
The availability of in vitro cell culture derived from local species provides opportunity for tackling problems related to preservation of its genetic materials and can potentially be applied for downstream in vitro-based studies. Here, we established primary fibroblast cell culture from Indonesian local chicken, Pelung chicken, then explored its growth characteristic and potential uses for wound healing assay and cytotoxicity tests of medicinal bioactive compounds. Fibroblast cells were isolated from embryonic skin tissue using explant technique. Cells were maintained in DMEM-FBS media at 37° C. In vitro wound healing assay was performed by creating a “scratch” in a cell monolayer, followed by capturing the images of migrating cells at regular intervals. Cell viability was measured using tryphan blue dye exclusion assay in various doses of extracts of Centella asiatica leaves. Cells outgrowth from the skin explant revealed typical morphology of fibroblast-like cells, showing spindle shaped cells forming monolayer which firmly adhered to the substrate. Early passage of Pelung fibroblast cells reached maximum growth at 7.95×104 cells/cm2 after 5 days with population doubling time (PDT) of 184 h. With continuous passage, population of the cells became more homogeneous and PDT was higher. In the wound healing assay, Pelung fibroblast cells migrated towards the wound area and after 24 hours closing the whole area, suggesting their ability to normally respond mechanical stimuli. In the cytotoxicity test, the viability of the cells corresponded in a dose-dependent manner with the amount of Centella asiatica extract tested into the culture.