A Method of Primary Cell Culture for Establishing Equine Long-Term Culture Cell Lines

Abstract

We investigated a method of culture for the establishment of equine long-term culture cell lines. The tissues tested were equine fetus tissues; right abdomen epithelium, the umbilical artery, the umbilical ring, the pulmonary artery, the renal cortex and the fetal placenta. To decide on a suitable primary culture method, we compared two methods of primary culture: tissue fragment culture and low-temperature trypsin cell-dispersal. On the other hand, to decide on a suitable culture medium, we compared 11 culture media: BME, MEM, D-MEM, IMDM and HFM (commercially completed culture media, available Human Foreskin Melanocyte). These were supplemented with 10% or 20% FCS when used as culture media. Moreover, HFM medium was used with no FCS, 10% and 20% FCS. Culture conditions were 37°C, 100% humidity, 5% CO₂ and 95% air. We obtained the following results: 1) The tissue fragment culture method was suitable for preparing primary cell culture from fetal tissue. 2) High-nutrition culture media such as D-MEM and IMDM were suitable for the culture of fetal tissues. 3) We established a cell line from cells of renal cortex, named “TEK-1”.