P-100: Determination of The Proper Pregnancy Age for Collection and Culture of Guinea Pig Fetal Fibroblast

Background: Fibroblast cell lines have different application in biological research including nuclear transfer in cloning, feeder layer of embryonic stem cells, wound healing research and tissue engineering. The objective of the present study was to determine the proper pregnancy age for collecting and culturing guinea pig fetal fibroblast. Materials and Methods: Guinea pig fetuses were collected after determination of their age (day 20, 30 and 45 of pregnancy) by measuring the fetal biparietal diameter using a real-time B-mode ultrasound scanner (SIUI V900, China) equipped with a 7.5 MHz, linear-array transducer. Skin of fetuses was cut into small pieces (1 mm2) and cultured in 88% Dulbecco's Modified Eagle Medium (DMEM), 10% fetal bovine serum (FBS), 1% penicillin and streptomycin, 1% L-glutamine in 25 cm2 flasks. When the cell cultures reached 80-90% confluence, they were passaged. The time intervals between the first and second passages in three different ages were determined.Results: The time interval between the first and second passages for guinea pig fetal fibroblasts were 30 days for day 20 pregnancy, 10 days for day 30 pregnancy, and 15 days for day 45 pregnancy. Conclusion: The proper pregnancy age for collection and culture of guinea pig fetal fibroblast was at the age of day 30 of pregnancy which made the shortest interval for reaching the maximum confluence in the fibroblast cell culture.