Following the successful growth of HBVPs in Poly-L-Lysine coated glass Petri dishes, I have enough of my own fibroid cells to repeat the process.
My cells continue to grow so slowly that I should be able to passage them into the Petri dishes and allow them to grow to confluence during the festive season break over 2 weeks . Of course, I need to clear this plan with Jo-Maree. No one else is using the incubator, so it should not be too much of a problem.
As part of this plan, I will be growing my tumour baby cells in 90mm glass Petri dishes and 1 x 90mm crystal dish. As per my previous experiment with HBVP cells, I need to coat the glass surface with Poly-L-Lysine solution to enable cell adherence.
I diluted the Poly-L-lysine solution with sterile MilliQ water (sterilised 14/12/21) to make up 40 mL total (10mL for each 900mm Petri dish x 3, plus 1 x cut glass crystal dish)
6mL PLL + 34mL MilliQ = 40mL PLL Solution
I added 10mL of the Poly-L-Lysine solution to each dish and then incubated them for an hour. [ The cut glass crystal dish was placed inside a 150mm autoclaved Petri dish to preserve sterility.]
Unwrapping Petri dishes and getting ready to coat culture glassware with PLL.
PLL coated glassware in Petri dishes ready for incubation.
Following incubation, I removed the Poly-L-Lysine solution and washed the dish with PBS. During cell passage of my confluent flask, I added 1mL of cell solution (from a 10mL suspension) and 5mL media. I placed the cut glass vessels back into a 150 mm Petri dish and into the incubator.
Cut glass vessels with cells ready for incubation.
Fingers crossed that they survive the holiday break!