4 CULTURE OF ANIMAL CELLS

encouraged toxicity and virological studies in insects, and
developments in gene technology have suggested that insect
cell lines with baculovirus and other vectors may be useful
producer cell lines because of the possibility of inserting
larger genomic sequences in the viral DNA and a reduced
risk of propagating human pathogenic viruses. Furthermore,
the economic importance of fish farming and the role of
freshwater and marine pollution have stimulated more studies
of normal development and pathogenesis in fish. Procedures
for handling nonmammalian cells have tended to follow those
developed for mammalian cell culture, although a limited
number of specialized media are now commercially available
for fish and insect cells (see Sections 27.7.1, 27.7.2).
The types of investigation that lend themselves
particularly to tissue culture are summarized in Fig. 1.2:
(1) intracellular activity, e.g., the replication and transcription
of deoxyribonucleic acid (DNA), protein synthesis, energy
metabolism, and drug metabolism; (2) intracellular flux, e.g.,
RNA, the translocation of hormone receptor complexes
and resultant signal transduction processes, and membrane
trafficking; (3) environmental interaction, e.g., nutrition,
infection, cytotoxicity, carcinogenesis, drug action, and
ligand–receptor interactions; (4) cell–cell interaction, e.g.,
morphogenesis, paracrine control, cell proliferation kinetics,
metabolic cooperation, cell adhesion and motility, matrix
interaction, and organotypic models for medical prostheses
and invasion; (5) genetics, including genome analysis in
normal and pathological conditions, genetic manipulation,
transformation, and immortalization; and (6) cell products
and secretion, biotechnology, bioreactor design, product
harvesting, and downstream processing.
The development of cell culture owed much to the
needs of two major branches of medical research: the
production of antiviral vaccines and the understanding of
neoplasia. The standardization of conditions and cell lines for
the production and assay of viruses undoubtedly provided
much impetus to the development of modern tissue culture
technology, particularly the production of large numbers
of cells suitable for biochemical analysis. This and other
technical improvements made possible by the commercial
supply of reliable media and sera and by the greater control of
contamination with antibiotics and clean-air equipment have
made tissue culture accessible to a wide range of interests.
An additional force of increasing weight from public
opinion has been the expression of concern by many animal-
rights groups over the unnecessary use of experimental
animals. Although most accept the idea that some
requirement for animals will continue for preclinical trials
of new pharmaceuticals, there is widespread concern that
extensive use of animals for cosmetics development and
similar activities may not be morally justifiable. Hence,
there is an ever-increasing lobby for more in vitro assays,
the adoption of which, however, still requires their proper
validation and general acceptance. Although this seemed a
distant prospect some years ago, the introduction of more
sensitive and more readily performed in vitro assays, together
with a very real prospect of assaying for inflammation in vitro,
has promoted an unprecedented expansion in in vitro testing
(see Section 22.4).
In addition to cancer research and virology, other areas
of research have come to depend heavily on tissue culture
techniques. The introduction of cell fusion techniques (see
Section 27.9) and genetic manipulation [Maniatis et al., 1978;
Sambrook et al., 1989; Ausubel et al., 1996] established
somatic cell genetics as a major component in the genetic
analysis of higher animals, including humans. A wide range
of techniques for genetic recombination now includes DNA
transfer [Ravid & Freshney, 1998], monochromsomal transfer

CELL PRODUCTS:
Proteomics, secretion,
biotechnology, biorector design,
product harvesting, down-
stream processing
INTRACELLULAR ACTIVITY: IMMUNOLOGY: Cell surface epitopes,
DNA transcription, protein synthesis, hybridomas, cytokines and signaling,
energy metabolism, drug metabolism, inflammation
cell cycle, differentiation, apoptosis
GENOMICS: Genetic analysis,
INTRACELLULAR transfection, infection,
FLUX: RNA processing, transformation, immortalization,
hormone receptors, senescence
metabolite flux, calcium
mobilization, signal
TISSUE ENGINEERING: Tissue
transduction, membrane
constructs, matrices and
trafficking
scaffolds, stem cell sources,
propagation, differentiation
PHARMACOLOGY: Drug
action, ligand receptor
interactions, drug metabolism, TOXICOLOGY: Infection,
drug resistance CELL-CELL INTERACTION: cytotoxicity, mutagenesis,
Morphogenesis, paracrine control, cell carcinogenesis, irritation,
proliferation kinetics, metabolic inflammation
cooperation, cell adhesion and motility,
matrix interaction, invasion

Fig. 1.2. Tissue Culture Applications.