Depending on the cell type used, scaffolds can be designed for a specific application and typically act as a template for bio-mimetic tissue. In order to achieve this, the tissue scaffold must be first vascularized, thereby allowing blood to freely move through the construct. Skin equivalents have been created by co-culturing dermis with epidermis with interviewing layers of collagen. Lastly, autoclave the scaffolds in a wet cycle for 20 minutes.
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Organotypic culture is defined as the culture of an organ collected from an organism. It is one method allowing the culture of complex tissues or organs. It allows the preservation of the architecture of the cultured organ and most of its cellular interactions.
Figure 1: Illustration of various organotypic culture method. Historical background While cells lines are one of the most often used experimental model by scientists, surprisingly, culture of whole organs start few decades before the first establishment of cell lines. Loeb managed to cultivated organs of rabbit. Quickly after, other authors report the culture of different organs collected from different organisms. In the meantime, the first establishment of cell lines has been done, slowing the development of organotypic culture.
The HeLa cell line marked the beginning of the intensive utilization of such a system by the scientific community. For example, HeLa cells are now cited in more than articles and part of patents. MCF-7 cells another classically used cell line are cited in more than articles. It reflects the intensive use of cell lines in research. Assets and drawbacks The massive use of cell line by researchers comes from the need to have a robust, reproducible, cheap and fast experimental model, that are key features of cell line cultures.
Despite all of these advantages, some drawbacks appear over time. Second is the loss of certain cell functions because the culture conditions are driven by the growth and not by the expression of a tissue-like phenotype. An illustration of this is the metabolism in the liver and liver cell lines. This bias makes the extrapolation of cell lines data to human tricky or even unrealistic. Figure 3: Schematic view of features for different culture model: from fractions to organotypic culture Since few years, the pressure of regulation agencies to improve the risk assessment for humans has lead to re-consider the use of organotypic cultures.
Whereas they are more expensive, more complex and time-consuming, they offer better extrapolation possibilities and are more relevant by taking into account the physiological environment. One of the best models is the culture of human organs, but the difficulties to handle such a system and to obtain such organs push researchers towards new systems offering both the advantages of cell cultures and organs culture. From organotypic culture to 3D models As mentioned above, there is now a need for developing new in vitro systems offering the advantages of both cell lines and organ culture.
One of the solutions is the 3D cell cultures , eg. The development of such a technique has to be based on the knowledge acquired with the two models. Figure 4: A: representation of the derivation of human cell cultures to obtain human spheroids ; B: synergy and inter-dependency of different cultures approaches.
From Pamies and Hartun, 21st century cell culture for 21st century toxicology, Chemical Researchin toxicology, Discover the CubiX.
Histotypic Tissue Culture
The organotypic model consists of three approaches for the original structural and functional interactive relationships of the organ. 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. These cellular interactions mostly due to hormonal stimulation are very important for the expression of their functions, as indicated by the following examples.
HISTOTYPIC CULTURE PDF
This video introduces histotypic tissue culture, where the growth and propagation of one cell line is done in an engineered three-dimensional matrix to reach high cell density. Here, we show the harvesting of cells from donor tissue, followed by cell culture on an engineered construct. Histotypic Tissue Culture. Histotypic tissue culture allows cells to be grown in three dimensions, thereby creating in-vitro tissue morphologies that closely mimic realistic tissue function, which can be used as viable constructs for tissue repair. These cultures are typically three dimensional structures consisting of a single cell type grown in high density.