title: An in vitro monocyte culture method and establishment of a human monocytic cell line (K63).
abstract: A novel method of monocyte culture in vitro was developed. The fraction of monocytes was obtained by density centrifugation of heparinised human venous blood samples. Monocytes were suspended in a modified Rosewell Park Memorial Institute medium (RPMI)-1640 (mRPMI) supplemented with 10% non-inactivated autologous serum added to the feeder cells. An avian cell line was used for feeder cells. Only those monocytes that settled on feeder cells grew rapidly at 37°C-38°C into a formation of clumped masses within two to three days. The cell mass was harvested and subcultures were made without feeder cells. A stable cell line (K63) was established from subcultures using a limited dilution method and cell cloning in microplates. K63 cells were adapted for later growth in the mRPMI medium supplemented with 10% foetal calf serum. The cells were well maintained at over 50th passage levels. This method proved to be applicable for monocyte cultures of animals as well.
authors.affiliation: Laboratory of Veterinary Microbiology, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-8510, Japan. katsuyukikadoi@yahoo.co.jp
authors.forename: Katsuyuki
authors.initials: K
authors.lastname: Kadoi
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D006801:Humans; D009000:Monocytes