Part I: Introduction to animal cell culture
Welcome to a new series of articles aimed at providing useful hints for
culturing animal cells (i.e., cells derived from higher eukaryotes such
as mammals, birds, and insects ). This article introduces different types of animal cell cultures.
The series will continue in future issues of QIAGEN News with considerations
for cell culture and cell culture protocols.
Animal cell cultures
Depending on their origin, animal cells grow either as an adherent monolayer
or in suspension.
are anchorage-dependent and propagate as a monolayer
attached to the cell culture vessel. This attachment is essential for proliferation
many adherent cell cultures will cease proliferating once they become
confluent (i.e., when they completely cover the surface of cell culture
vessel), and some will die if they are left in this confluent state for
too long. Most cells derived from tissues are anchorage-dependent.
can survive and proliferate without being attached
to a substratum. Hematopoietic cells (derived from blood, spleen, or bone
marrow) as well as some transformed cell lines and cells derived from malignant
tumors can be grown in suspension.
, finite cultures, and continuous cell lines differ
in their proliferative potential (see below). Different cell types vary
greatly with respect to their growth behavior and nutritional requirements.
Optimization of cell culture conditions is necessary to ensure that cells
are healthy and in optimal condition for downstream applications.
Tip- Extensive informat
ion on culturing cells can be found in reference
Primary cell cultures
Primary cell cultures come from the outgrowth of migrating cells from a
piece of tissue or from tissue that is disaggregated by enzymatic, chemical,
or mechanical methods. Primary cultures are formed from cells that survive
the disaggregation process, attach to the cell culture vessel (or survive
in suspension), and proliferate.
Primary cells are morphologically similar to the parent tissue. These cultures
are capable of only a limited number of cell divisions, after which they
enter a nonproliferative state called senescence and eventually die out.
Adherent primary cells are particularly susceptible to contact inhibition,
that is, they will stop growing when they have reached confluency. At lower
cell densities, however, the normal phenotype can be maintained. Primary
cell culture is generally more difficult than culture of continuous cell
Primary cell cultures are sometimes preferred over continuous cell lines
in experimental systems. Primary cells are considered by many researchers
to be more physiologically similar to in vivo cells. In addition, cell lines
cultured for extended periods of time can undergo phenotypic and genotypic
changes that can lead to discrepancies when comparing results from different
laboratories using the same cell line. Furthermore, many cell types are
not available as continuous cell lines.
Finite cell cultures
Finite cell cultures are formed after the first subculturing (passaging)
of a primary cell culture. These cultures will proliferate for a limited
number of cell divisions, after which they will senesce. The pro
potential of some human finite cell cultures can be extended by introduction
of viral transforming genes (e.g., the SV40 transforming-antigen genes).
The phenotype of these cultures is intermediate between finite cultures
and continuous cultures. The cells will proliferate for an extended time, but usually the culture will eventually cease dividing,
similar to senescent primary cells. Use of such cells is sometimes easier
than use of primary cell cultures, especially for generation of stably transfected
Continuous cell lines
Finite cell cultures will eventually either die out or acquire a stable,
heritable mutation that gives rise to a continuous cell line that is capable
of unlimited proliferative potential. This alteration is commonly known
as in vitro transformation or immortalization and frequently correlates
Rodent primary cell cultures form continuous cell lines relatively easily,
either spontaneously or following exposure to a mutagenic agent. In contrast,
human primary cell cultures rarely, if ever, become immortal in this way
and require additional genetic manipulation to form a continuous cell line.
However, cell cultures derived from human tumors are often immortal.
Continuous cell lines are generally easier to work with than primary or
finite cell cultures. However, it should be remembered that these cells
have undergone genetic alterations and their behavior in vitro may not represent
the in vivo situation.
The QIAGEN Guide to Animal Cell Culture will continue in future issues of
QIAGEN News with tips for successful cell culture. If there is any other
information you would like to see on these pages of QI
AGEN News, please
let us know by calling QIAGEN Technical Services or your local distributor.
1. Freshney, R.I. (1993)
Culture of Animal Cells,
A Manual of Basic Technique,
3rd ed., New York:
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. The QIAGEN Guide to Animal Cell Culture2
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