ADVANCED TOOLS FOR TISSUE ENGINEERING, BIOMATERIAL TESTING AND 3D CULTURES

The advantages:

In case you want to multiply cells, do not read any further information, because perfect conventional methods are available for your purpose.

However, if you want to generate functional, thus highly differentiated tissues then the following new information may be the basis for your future work.

The per(i)fusion culture system presented here makes experiments possible, which failed by conventional methods.

Advantages are:

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Selection from a variety of cell supports
Individual influence on differentiation
Per(i)fusion of culture medium
Culture of epithelia in a natural gradient
Microscopical observation
Long term culture for chronical toxicity
Tissue engineering for surgical implantation
Testing of biomaterials

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We show a new strategy to generate differentiated tissues. All of our tools for tissue engineering were designed and constructed arising out of experimental necessities and professional experience.

Culture

Culture conditions as close as possible to the situation within an organ:

The value of cultured cells and tissues in biomedical research is highly dependent on their degree of terminal differentiation. In the stagnant environment of conventional culture dishes however, cells and tissues lose their morphological, physiological and biochemical features by dedifferentiation. As a result they are not comparable to the situation within in an organ any more. Because many experiments failed in conventional dishes, we developed a new technique to obtain improved differentiation.

In optimally cultured tissue. This was the reason why many researchers started asking for the system. The increasing demand for our tools finally was the reason for Katharina Lorenz-Minuth to found a private company in 1993.

We are proud that our project was awarded by the Philip Morris research prize “Challenge of the future” in 1992.
MINUSHEET is a registered trade mark.

From numerous experiments we know that optimal differentiation can only be obtained, if the environment is adapted as close to the natural situation as possible. Furthermore we experienced that differentiation of cells and tissues in vitro depends not only on a single morphogenic substance but on several environmental factors.
Cells and tissues are kept on interchangeable supports mounted in special carriers, the MINUSHEEETS. This way an ideal support for the cells can be selected. Optimal support and cell adhesion is an important presuppostion for differentiation.

The developed per(i)fusion culture system is based on the following patents: DE 39 23 279,

DE 42 00 446,
DE 42 08 805,
DE 44 43 902,
DE 19 530 556,
DE 196 48 876 C2,
US 5 190 878,
US 5 316 945,
US 5 665 599,
J 2847669,
PA 10 2005 001 747.9,
PA 10 2004 054 125.6,
PA 10 2005 002 938.8,

and others. For ecological reasons all of the parts can be autoclaved and are constructed for repeated use.
The cell and tissue carriers can be used in conventional tissue culture plates or they are placed in newly constructed per(i)fusion culture containers. A continuous flow of culture medium guarantees optimal nutrition, prevents an overshoot of paracrine factors and keeps harmfull metabolic products on a constantly low level.
The advanced culture system was developed by Will W. Minuth and his team at the Department of Anatomy, University of Regensburg, Regensburg, Germany. Part of the cell biological research was supported by the Deutsche Forschungsgemeinschaft (DFG). The consequent experimental work resulted.

Selecting a suitable support for optimal cell anchorage and differentiation:

Cell and tissue supports are necessary for advanced culture experiments. Numerous materials, for example different kind of filters are commercially available in diamters of 13 and 47 mm from:
Millipore
Gelman Sciences
Nuclepore
Nunc
Schleicher Schüll
Sartorius
Whatmann
and a large variety of other companies.

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Excellent culture experiences are made with:

Glass cover slips
Whatman transparent
Melafol
Cellophane
Thermanox
Polycarbonate Microclear
Nylon 80555
Polycarbonate black
Polycarbonate white
Polycarbonate HTTP
Nuclepore
Microclear Polycarbonate
Cellulosenitrate HAWP
Capsula fibrosa from kidney

All kinds of cells can also be easily recognized on non-transparent supports after labeling them with fluorescent markers such as DAPI or propidium iodide.