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MINUCELLS AND MINUTISSUE VERTRIEBS GMBH

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PAGE 13

Instructions for use part 2:


 

The tissue carriers consist of 3 parts: The black base ring (1), the selected support material for optimal cell attachment and differentiation (2) and the small white tension ring (3). For mounting the support material is placed into the black base ring (4), then the white tension ring is snapped into the base ring (5). Mounting of the tissue carrier is completed.

Dismantling a MINUSHEET tissue carrier: All of the newly constructed cell culture devices are constructed for repeated use. If the support material within a MINUSHEET has to be changed, the following procedure is recommended. Use the tip of a fine forceps (6,7) and lift the tension ring from the inner side (8). Remove the ring and exchange the support material.

Sterilization: Depending of the support material the tissue carriers are placed into a Melafol envelope, a metal box or a glass dish. The tissue carriers can be briefly sterilized in solutions such as ethanol and formaline.

Limits for sterilization: The optimum is autoclaving at 105°C and 0.3 bar for 20 minutes, the absolute maximum is 135°C and 2 bar for 10 minutes.
Do not close the stainless clamps of the culture container during the autoclaving procedure!

Medium is added with a small pipette into the cleft between the base of the culture plate and the tissue carrier until the support is wet. Then the cells are pipetted onto the surface of the support in a small volume. In order for the cells to adhere to the support, the dish is transferred to an incubator. Mostly within a few hours the cells are attached and are ready for further experimenting. Tissue slices can be placed between two nylon nets in the tissue carrier as a sandwich.






Assembly of a working line for per(i)fusion culture
Autoclaving of MINUSHEETS

Preculture of cells: For a culture experiment the sterilized tissue carriers are transferred into a glass dish or a 24 well culture plate with fine forceps.

Assembly of a working line for
per(i)fusion culture:

A working line consists of 2 glass bottles (10), 2 screw caps (11), a cell culture container (12), silicone tubes with luer fittings (13). First, the screw caps are mounted on the bottles (14), then the culture container is connected to the screw caps through the silicone tubes. Inlet at the base, outlet at the top side of the container (15). For sterilization the assembled working line is autoclaved with the lid of the culture container closed but not clamped down.


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