7
6. Make sure that the water pump is pumping water from the reservoir and through
the Control and Stretch Unit.
7. Turn off Main Power and Water Pump Switches.
►
Start Cell Stretching
1. The Power Switch must be OFF to freely rotate the Chamber Length
Adjustment Knob.
Silicone chambers containing cultured cells are attached to the
Stretch Unit by inserting the 4 pins into the corners of the chamber. Rotate the
Adjustment Knob clockwise until the bottom of the chamber is taut.
2. Select the stretch parameters by changing the Stretch Pattern Buttons. Refer to
Section 3 for stretch parameters.
3. Do not change the Strain Ratio or Strain Frequency Selector Buttons
during the operation of a stretch cycle. Press the Stop Button and wait
for the last cycle to complete and the chamber returns to its starting
position before changing stretch parameters.
Changing parameters during
a stretch cycle may damage the motors.
4. Push the Start Button to start the stretch unit.
5. After 5 minutes, stop the stretching and check the condition of the cells. If the cells
have not detached, proceed with your experiment. If the cells are detached, the
adhesion matrix coating was probably insufficient. Recoat the chambers.
Culturing Cells in the Silicone Chambers
1. Seed cells at the appropriate concentrations in the freshly coated chambers.
Important:
It is critical to not over expose the cells to dissociation enzymes. Cells
should be treated in the same manner (type, concentration of enzyme,
temperature, and exposure time) for all experiments.
Important:
Cells should not be cultured at a high cell density in the chambers. For
example, epithelial cells often form a cell-sheet and the cell-cell adhesion seems
to be stronger than a cell-surface adhesion. When this happens cells may detach
from the chamber. Additionally, cultures that are grown over a week in the
chambers may detach.
2. After overnight incubation, inspect cells under a microscope to ensure that they
adhered to the chamber.
