Describe the invasion assay using matrigel
- Can draw
- Assay to test invasiveness of a cell line
- Matrigel is mimicking the ECM
- Is in a cell culture insert and media is bathing ontop of cell
- If they are invasive they are neoplastic cells.
Long term storage done with cryopreservation and uses cryovials. Small vials.
Cryopreservation = ultrafreezin temp in liquid N.
Cryoprotectants protectant during freezing time, otherwise you get cell damage.
Prokaryotic cells lot more resistant to freezing than mammalian cells.
Coolin rate 1 degree per minute. Want it to slowly cool so u don’t get ice crystals and then warm it really
Mycoplasma contamination very difficult to deal with
- Are horrible, cant see them, very small, can damage cell culture and how they grow.
- Known as PPLO
- Smallest free living cells
- NO CELL WALL
o Many of the antibiotics target cell wall and that wont help with this
Source of this:
- Human origin
o One of the hoods is horizontal hood, perhaps u coughed and the mycoplasma could
have been introduced
o Sera = FBS contamination
o Trypsin = comes from pigs?
Number of ways u can test for it
- Can culture it, stain it