Basic Cell Culture Protocols

Basic Cell Culture Protocols 
THIRD EDITION
Edited by
Cheryl D. Helgason
Cindy L. Miller

Culture of Primary Adherent Cells

and a Continuously Growing Nonadherent Cell Line

Cheryl D. Helgason

Summary

Cell culture is an invaluable tool for investigators in numerous fields. It facilitates

analysis of biological properties and processes that are not readily accessible at the level

of the intact organism. Successful maintenance of cells in culture, whether primary or

immortalized, requires knowledge and practice of a few essential techniques. The purpose

of this chapter is to explain the basic principles of cell culture using the maintenance

of a nonadherent cell line, the P815 mouse mastocytoma cell line, and the isolation

and culture of adherent primary mouse embryonic fibroblasts (MEFs) as examples.

Procedures for thawing, culture, determination of cell numbers and viability, and

cryopreservation are described.