HOME

TheInfoList



OR:

Primary cell culture is the ''
ex vivo refers to biological studies involving tissues, organs, or cells maintained outside their native organism under controlled laboratory conditions. By carefully managing factors such as temperature, oxygenation, nutrient delivery, and perfusi ...
''
culture Culture ( ) is a concept that encompasses the social behavior, institutions, and Social norm, norms found in human societies, as well as the knowledge, beliefs, arts, laws, Social norm, customs, capabilities, Attitude (psychology), attitudes ...
of cells freshly obtained from a multicellular organism, as opposed to the culture of immortalized cell lines. In general, primary cell cultures are considered more representative of ''
in vivo Studies that are ''in vivo'' (Latin for "within the living"; often not italicized in English) are those in which the effects of various biological entities are tested on whole, living organisms or cells, usually animals, including humans, an ...
'' tissues than cell lines, and this is recognized legally in some countries such as the UK ( Human Tissue Act 2004). However, primary cells require adequate substrate and nutrient conditions to thrive and after a certain number of divisions they acquire a senescent phenotype, leading to irreversible
cell cycle The cell cycle, or cell-division cycle, is the sequential series of events that take place in a cell (biology), cell that causes it to divide into two daughter cells. These events include the growth of the cell, duplication of its DNA (DNA re ...
arrest. The generation of cell lines stems from these two reasons. Primary cells can become immortalized either spontaneously (e.g.
HeLa cells HeLa () is an immortalized cell line used in scientific research. It is the oldest human cell line and one of the most commonly used. HeLa cells are durable and prolific, allowing for extensive applications in scientific study. The line is ...
) or by genetic modification (e.g. HEK cells), at which point they become cell lines which can be subcultured indefinitely. Because of their requirements for viability, primary cell cultures did not become widespread until the 2000s. These cultures present several advantages over cell lines, including a better representation of the cellular heterogeneity of tissues, a more faithful transcriptomic and proteomic profile (especially when cultured in 3D) and more realistic functional responses, including drug responses. In contrast, immortalized cell lines are known to become homogeneous through the natural selection of specific subpopulations, to undergo
genetic drift Genetic drift, also known as random genetic drift, allelic drift or the Wright effect, is the change in the Allele frequency, frequency of an existing gene variant (allele) in a population due to random chance. Genetic drift may cause gene va ...
and to acquire genetic aberrations. In many cases, cell lines have been misidentified, contaminated with other cells or infected with ''
Mycoplasma ''Mycoplasma'' is a genus of bacteria that, like the other members of the class ''Mollicutes'', lack a cell wall, and its peptidoglycan, around their cell membrane. The absence of peptidoglycan makes them naturally resistant to antibiotics ...
'', small intracellular
bacteria Bacteria (; : bacterium) are ubiquitous, mostly free-living organisms often consisting of one Cell (biology), biological cell. They constitute a large domain (biology), domain of Prokaryote, prokaryotic microorganisms. Typically a few micr ...
that went undetected for decades. When whole or partial tissues are isolated and maintained ''ex vivo'', the procedure is termed primary
tissue culture Tissue culture is the growth of tissue (biology), tissues or cell (biology), cells in an artificial medium separate from the parent organism. This technique is also called micropropagation. This is typically facilitated via use of a liquid, semi-s ...
. More specific terms include organotypic culture, tissue slices and explants. Neuronal primary cell cultures are cells collected from the brain of an organism. For example they can be used when examining substances effect on cell viability, which can further on be potential treatments for brain deficits.


Monolayer cultures

Monolayer cultures refer to
cell culture Cell culture or tissue culture is the process by which cell (biology), cells are grown under controlled conditions, generally outside of their natural environment. After cells of interest have been Cell isolation, isolated from living tissue, ...
s where cells are grown in a single, flat layer on the surface of a
culture dish A Petri dish (alternatively known as a Petri plate or cell-culture dish) is a shallow transparent lidded dish that biologists use to hold growth medium in which cells can be cultured,R. C. Dubey (2014): ''A Textbook Of Biotechnology For Class-X ...
or substrate. In a monolayer culture, cells adhere to the substrate and spread out in a two-dimensional arrangement. This type of cell culture is commonly used in
laboratory A laboratory (; ; colloquially lab) is a facility that provides controlled conditions in which scientific or technological research, experiments, and measurement may be performed. Laboratories are found in a variety of settings such as schools ...
settings for various purposes, including research, drug testing, and biotechnology. Key features of monolayer cultures include: * Two-Dimensional Growth: Cells in monolayer cultures grow in a single plane, adhering to the surface of the culture vessel. This flat arrangement allows for easy observation and manipulation of individual cells. *Adherence to Substrate: The cells attach to the surface of the culture dish or flask, and their growth and behavior can be influenced by the characteristics of the substrate.on the other words In cell culture, adherence to substrate describes a cell's capacity to adhere to a surface and proliferate. Many elements, including surface energy, substrate topography, and roughness, mediate the process of cell attachment. The study of artificial polymer surfaces with varying chemical, topological, and mechanical cues that regulate cell activities has focused attention on the interaction between external surfaces and cells. In a study that was published in the journal RSC Advances in 2021, the impact of roughness and surface energy on cell adhesion and growth was examined. The most advantageous circumstances for effective cell adhesion, development, and proliferation were discovered by the study to be moderate surface energy and intermediate roughness ratio. * Cell Proliferation: Cells in monolayer cultures can undergo cell division and proliferation. This feature is crucial for experimental studies and the production of a larger number of cells for subsequent analyses. * Observation and Imaging: The two-dimensional nature of monolayer cultures makes it convenient for microscopic observation and imaging. Researchers can easily visualize the cells, study their morphology, and monitor changes over time. * Cell Differentiation: Depending on the cell type and culture conditions, monolayer cultures can be used to induce cell differentiation. This is particularly important in studying developmental processes and tissue-specific functions.


Monolayer cultures for personalized therapy

The endocrine cancer with the highest incidence is
thyroid cancer Thyroid cancer is cancer that develops from the tissues of the thyroid gland. It is a disease in which cells grow abnormally and have the potential to spread to other parts of the body. Symptoms can include swelling or a lump in the neck, ...
(TC). Differentiated thyroid cells (DTC) that originate from follicular thyroid cells account for over 90% of total thyroid cells (TC). Papillary TC (PTC), follicular TC (FTC), and Hürthle cell TC are examples of DTC. One percent of TC is anaplastic TC (ATC), which accounts for 15–40% of TC deaths. Mortality is one of the biggest obstacles to current treatment techniques against aggressive DTC or ATC. These strategies are not entirely effective against these conditions. Recent years have seen advancements in our knowledge of the molecular and genetic underpinnings of TC development as well as the introduction of novel medications, such as
tyrosine kinase inhibitors A tyrosine kinase inhibitor (TKI) is a pharmaceutical drug that inhibits tyrosine kinases. Tyrosine kinases are enzymes responsible for the activation of many proteins by signal transduction cascades. The proteins are activated by adding a phosph ...
(TKIs), which target the oncogenic or signaling kinases linked to cellular proliferation. Preclinical models have made use of thyroid cell lines that were isolated from tumor cells and selected for their high rate of proliferation in vitro. As a result of their adaptation to in vitro growth circumstances, these cells actually lose the distinctive characteristics of the original tumor. Because of these factors, there are significant restrictions on the usage of these cell lines. More recently, monolayer cultures of human primary cells have been created, and their biological behavior has been studied. Furthermore, whereas human primary cell cultures may now be created from samples of fine-needle aspiration cytology from aggressive dedifferentiated DTC or ATC, primary TC cells were previously only obtained by surgical biopsies. Without the use of useless medications, testing several TKIs in vitro on individual patients can aid in the development of novel, individualized treatments. Limitations of monolayer culture and motivations: Scientists are investigating novel models that can more accurately mimic the structure and function of human organs because to the limitations of monolayer culture settings. Protocol improvements in recent times have led to the creation of three-dimensional (3D) organ-like architectures known as "organoids," which are able to exhibit properties of their corresponding real organs, such as morphological features, functional activities, and individual responses to particular
pathogens In biology, a pathogen (, "suffering", "passion" and , "producer of"), in the oldest and broadest sense, is any organism or agent that can produce disease. A pathogen may also be referred to as an infectious agent, or simply a germ. The term ...
.


Cell culture protocol

For in vitro investigations to be conducted correctly, the cell culture protocol for a particular cell line must be optimized. The best culture conditions for different cell lines can differ significantly due to the
heterogeneity Homogeneity and heterogeneity are concepts relating to the uniformity of a substance, process or image. A homogeneous feature is uniform in composition or character (i.e., color, shape, size, weight, height, distribution, texture, language, i ...
of germ cell malignancies.


See also

* 3D cell culture *
Induced pluripotent stem cell Induced pluripotent stem cells (also known as iPS cells or iPSCs) are a type of pluripotent stem cell that can be generated directly from a somatic cell. The iPSC technology was pioneered by Shinya Yamanaka and Kazutoshi Takahashi in Kyoto, Jap ...
* Patient-derived xenograft * Stem cell laws


References

{{reflist, 2 Cell culture