In vitro fibroblast cells culture from Pelung chicken embryo and its potential application

Abstract

Abstract The availability of in vitro cell culture derived from local breed provides an opportunity for tackling problems related to the preservation of its genetic materials and can potentially be applied for downstream in vitro-based studies. Here, we established primary fibroblast cell culture from 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 and maintained in DMEM-FBS media. Wound healing assay was performed by creating a “scratch” in the cell monolayer, followed by capturing periodic images of migrating cells. Cell viability was measured using trypan blue dye exclusion assay in various doses of Centella asiatica L. leaf extract. Cells outgrowth from the skin explant revealed a typical morphology of fibroblast-like cells that reached maximum growth at 7.95 × 104 cells/cm2 after 5 days. With continuous passage, the population of the cells became more homogeneous and population doubling time increased. In the wound healing assay, cells migrated towards the wound area within 24 hours, suggesting their ability to normally respond to chemical cues. In cytotoxicity test, cells’ viability corresponded in a dose-dependent manner with the amount of C. asiatica extract tested into the culture.