F
LEXCELL
®
I
NTERNATIONAL
C
ORPORATION
22
Cells can be lysed with appropriate buffer
to collect total RNA or intracellular
proteins.
The Culture Slips
®
can be returned to their
original culture vessel for further incubation
and subsequent collection of cell
supernatant. The medium can then be
assayed for released effector molecules.
The cells can be trypsinized for replating or
counting.
APPLICATION NOTES
C
ULTURING
C
ELLS ON
C
ULTURE
S
LIPS
®
Culture Slips
®
are Teflon
®
-bordered 75 x 25 x 1 mm glass culture surfaces that are either untreated or
bonded with peptides of collagen, elastin, fibronectin (RGD repeat as Pronectin F), laminin (as the
YIGSR peptide). The Teflon
®
border provides a means to culture cells only in the flow area. Bonded
peptides increase cell attachment.
Cells are plated on the growth surface at 10-25,000 cells/cm
2
in 3 to 5 ml of medium.
Be sure to plate
cells on the side where the Teflon
®
border is printed.
Once the cells are attached, additional medium
is added and the culture vessel placed into a CO
2
incubator at 37
°
C. Once the cells have grown to
confluence (normally 48 hours), the Culture Slips
®
are removed and inserted into the Streamer
®
flow
device for the experiment. Once the flow experiment is over, the Culture Slips
®
can be returned to their
original culture vessel to allow the measurement of secreted molecules post-flow.
If you experience cell detachment problems during flow regimes, try the following protocol for better
cell attachment to the Culture Slips
®
.
1.
Plate ½ of the normal amount of cells on the Culture Slips
®
.
2.
Reduce the media serum concentration (5% preferably) to slow the cell growth rate. This will
give the cells time to make their own protein matrix which will improve attachment.
3.
Allow the cells to grow to near confluency (4-5 days).
