Recently we established three fibroblast cell lines from ear skin explants of normal healthy dairy goats, of Kiko and Saanen breed (In Vitro Cell.Dev.Biol.-Animal; doi: 10.1007/s11626-010-9373-4). The cell lines revealed a viability rate of 96.2%, displayed a typical ‘S’ shaped growth curve with a population doubling time of 25 hours, and were free from microbial, fungal and mycoplasma contamination. We further characterized these cell lines and, in this communication we show the cytogenetic analysis and the genetic transfection of GSF289 cells. The GSF289 cell line which originated from Saanen breed of goats was successfully transfected with pcDNA3.1/NTGFP plasmid vector containing green fluorescent protein (GFP) gene under human cytomegalovirus (CMV) promoter. The efficiency of... transfection, as measured by flow cytometry, was 14.5% after 4 days of culture. The cytogenetic analysis performed on 29 G-banded metaphase cells revealed that the cell line has a normal male goat karyotype consisting of 58 autosomes and two XY sex chromosomes. These results suggest that GSF289 cell line with a normal karyotype, having a high rate of proliferation, and its ability to be easily transfected with plasmid DNA vectors is an additional tool to study molecular mechanisms that regulate fibroblast function as well as genetic manipulation of small ruminants.