Growth kinetics, plasticity and characterization of hamster embryonic fibroblast cells

Abstract

Development of different embryonic stem cells (ESCs) requires feeder fibroblast cell culture. Moreover, the establishment of fibroblast cell culture especially for endangered species can provide an excellent resource for biological research and preserve precious genetic materials. This study aimed to characterize and determine the growth kinetic of hamster embryonic fibroblast cells. Hamster fetuses of two albino female hamsters were collected between 8 and 10 days of pregnancy. After removal of the head, liver and gut, the fetuses were cut into 1-mm2 pieces and then cultured. After reaching 80–90% confluence, the cells were subcultured. The cells of passage 8 were subcultured in two 24-well plates (2 × 104 cells/well) for 7 days. Three wells per day were counted and the average cell counts at each time point were plotted against time, and the population doubling time (PDT) was determined. Cell viability after freezing and thawing was evaluated. For karyotyping, the cells of the passage 8 were used. The PDT of the cells at passage 8 was about 34.9 h and the viability was 77.9% in the passage 8. The isolated cells were spindle shaped and plastic adherent. The chromosome number and morphology were normal. The favorable morphology, viability, growth kinetic and karyotyping of hamster embryonic fibroblast cells revealed that these cells even at the eighth passage can safely be used as a feeder layer for ESCs, transgenic purposes and gene banks, and also for biological and pharmacological research.