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Lecture 9

BIOL483- Lecture 9 - Sept 28.docx

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BIOL 483
Vivian Dayeh

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 quickly. 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
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