Protocol

Inoculation and Passaging of Mammalian Monolayer Cell Cultures

This protocol was adapted from "Growth and Manipulation of Cells in Culture," Chapter 2, in Cells (eds. Spector et al.). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1998.

INTRODUCTION

Cells grown in monolayer proliferate to a confluent state in which the cells cover the growth surface of the flask. Some cells can be maintained in this plateau phase of growth for days to weeks, while others require trypsinization and subculture to survive. Cell lines requiring trypsinization usually do not display contact inhibition of growth and continue to proliferate. Ultimately, they peel off the surface of the flasks and are difficult to disperse and replate. This protocol describes a procedure for the subculture of mammalian cells in monolayer culture.