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The organotypic model consists of three approaches for the original structural
and functional interactive relationships of the organ.
The three approaches are: (1) Organ Cultures (2) Histotypic Cultures and (3)
Organotypic Cultures.
The cell cultures are widely used in the laboratories world over for various
purposes. In vitro studies with isolated cells are useful for understanding of many
cell functions such as transcription, translation, cell proliferation, respiration and
glycolysis. Thus for the study of biology and many functions, the cells grown in
conventional and monolayer cultures may be adequate.
content is associated with the risk of O2induced toxicity e.g. nutrient metabolite
exchange is severely affected.
Growth and Differentiation:
In general, the organ cultures do not grow except some amount of proliferation
that may occur on the outer cell layers.
In recent years, filter-well inserts are in use to attain the natural geometry of
tissues more easily.
2. Histotypic Cultures:
Growth and propagation of cell lines in three- dimensional matrix to high cell
density represent histotypic cultures. The advantage with this culture system is
that dispersed monolayer cultures can be used to regenerate tissue-like
structures. The commonly used techniques in histotypic cultures use gel and
sponge hollow fibers and spheroids.
Many workers claim that the behaviour of high-density cells formed on hollow
fibers is comparable to their in vivo behaviour. For instance, choriocarcinoma
cells grown in hollow fiber cultures release more chorionic gonadotrophin than in
a conventional monolayer. Hollow fiber culture techniques are regarded as ideal
systems for the industrial production of several biologically important
compounds. Work is progressing in this direction.
3. Organotypic Cultures:
Organotypic culture basically involves the combination of cells from different
lineages in a determined ratio to create a component of an organ. With the
advances in the organotypic culture techniques, it is now possible to develop
certain tissues or tissue models.
i. Skin equivalents have been created by co-culturing dermis with epidermis with
interviewing layers of collagen.
Cells that are cultured in suspension can be maintained in culture flasks that are not tissue-culture treated,
but as the culture volume to surface area is increased beyond which adequate gas exchange is hindered
(usually 0.2 0.5 mL/cm2), the medium requires agitation. This agitation is usually achieved with a
magnetic stirrer or rotating spinner flasks.
Requires periodic passaging, but allows Easier to passage, but requires daily cell
easy visual inspection under inverted counts and viability determination to
microscope follow growth patterns; culture can be
diluted to stimulate growth
Used for cytology, harvesting products Used for bulk protein production, batch
continuously, and many research harvesting, and many research
applications applications
http://www.biotechnology4u.com/animal_biotechnology_types_cell_cultures.html
The maintenance of growth of cells dissociated from the parental tissue (such as kidney, liver) using the
mechanical or enzymatic methods, in culture medium using suitable glass or plastic containers is called Primary
Cell Culture.
The primary cell culture could be of two types depending upon the kind of cells in culture.
a) Anchorage Dependent /Adherent cells- Cells shown to require attachment for growth are set to be
Anchorage Dependent cells. The Adherent cells are usually derived from tissues of organs such as kidney where
they are immobile and embedded in connective tissue. They grow adhering to the cell culture.
b) Suspension Culture/Anchorage Independent cells - Cells which do not require attachment for growth or do
not attach to the surface of the culture vessels are anchorage independent cells/suspension cells. All suspension
cultures are derived from cells of the blood system because these cells are also suspended in plasma in vitro e.g.
lymphocytes.
When a primary culture is sub-cultured, it becomes known as secondary culture or cell line. Subculture (or
passage) refers to the transfer of cells from one culture vessel to another culture vessel.
Subculturing- Subculturing or splitting cells is required to periodically provide fresh nutrients and growing space for
continuously growing cell lines. The process involves removing the growth media, washing the plate, disassociating
the adhered cells, usually enzymatically. Such cultures may be called secondary cultures.
Cell Line
A Cell Line or Cell Strain may be finite or continuous depending upon whether it has limited culture life span or it is
immortal in culture. On the basis of the life span of culture, the cell lines are categorized into two types:
a) Finite cell Lines - The cell lines which have a limited life span and go through a limited number of cell
generations (usually 20-80 population doublings) are known as Finite cell lines. These cell lines exhibit the property
of contact inhibition, density limitation and anchorage dependence. The growth rate is slow and doubling time is
around 24-96 hours.
b) Continuous Cell Lines - Cell lines transformed under laboratory conditions or in vitro culture conditions give
rise to continuous cell lines. The cell lines show the property of ploidy (aneupliody or heteroploidy), absence of
contact inhibition and anchorage dependence. They grow in monolayer or suspension form. The growth rate is
rapid and doubling time is 12-24 hours.
c) Monolayer cultures - When the bottom of the culture vessel is covered with a continuous layer of cells, usually
one cell in thickness, they are referred to as monolayer cultures.
d) Suspension cultures - Majority of continuous cell lines grow as monolayers. Some of the cells which are non-
adhesive e.g. cells of leukemia or certain cells which can be mechanically kept in suspension, can be propagated in
suspension. There are certain advantages in propagation of cells by suspension culture method.