Establishing Bovine Epithelial Cells by Removing Fibroblast During Primary Culture

다국어 초록 (Multilingual Abstract)

Fibroblasts are usually mixed into primary cultures of epithelial cells during enzymatic digestion of animal tissues or organs. They proliferate quickly and grow over the population of epithelial cells. In order to establish epithelial cell culture, we investigated possible ways to eliminate fibroblasts using three epithelial-derived cultures of bovine fetus. After digestion of fetal tissues with collagenase and dispase, cells were cultured in the medium of Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) for 24 hours. The cultured cells were treated with 0.01% edetate disodium dihydrate (Na₂EDDA) and detached fibroblasts were removed by changing media. The epithelial cells with their own shape were confirmed under a microscope. If fibroblast like cells were still present after the first elimination, they were removed again with Na₂EDDA treatment after culturing cells in DMEM containing 2% FBS. This procedure was repeated until almost pure primary epithelial cultures were established. The optimal method of fibroblast elimination was treating cells with Na₂EDDA for 3 minutes, 24 hours after primary cultivation in 10% FBS media and followed by treating cells with Na₂EDDA for 2 minutes, 6 hours after cultivation in 2% FBS media. We were able to obtain almost pure epithelial cells by the additional treatment with Na₂EDDA for 3 minutes, 12-24 hours after cultivation in 2% FBS media.

목차 (Table of Contents)

• 재료 및 방법
• 결과
• 고찰
• 감사의 글
• 참고문헌