Facts Flashcards
(35 cards)
Alamar Blue colour change?
Blue to Red (Res to Res)
5 year myeloma survival?
47%
How could WT vs T proliferation rates have diverged?
- There was a period of work up (15x passages long), faster cells will have out competed slower growing cells
- JJN3s grow faster therefore a disruptive gene insertion is more likely, due to lentivirus slight preference for actively transcription areas.
- Fast growing JJN3-T may have expressed less GFP and been sorted to the waste basket
What percentage of MCs express CXCR4
~50%
What percentage of MCs express N-Cadherin
~50%
What percentage of MCs express CD162
~100%
What is the name of the sialyted version of CD162?
HECA452
Sialyted clones, better or worse prognosis?
Worse
Did I say I believe in U266 N-Cadherin expression or not?
I said I did believe in it
Did I say I believe in U266 N-Cadherin expression or not?
I said I did believe in it
Name the significant differences between WT vs T for membrane molecule expression
All for N-Cadherin, for CD162; OPM2 and U266
How could U266 N-Cadherin expression be validated?
Select and proliferate up, or immunohistochemistry
I said Lentivirus transduction is stable, therefore…
spontaneous non expression of GFP is not likely
Why do GFP cells have a long negative tail for adhesion molecule expression?
Unclear, but JJN3s have a clear negative population for N-Cadherin
Why are there differences between JJN3 WT vs T N-Cadherin expression?
- Possibly lentivirus integration near to N-Cadherin gene locus could have affected expression.
- Faster proliferating cells might spend less time expressing N-Cadherin, and have out competed
Crystal violet should have been more reliable due to homogenisation, therefore every cell should contribute to reading
A sample of the well bottom, only the cells in view contribute
Mechanism of PLL adhesion?
Ionic, relies on negative membrane potential maintained by all live cells
What do MG63 cells express?
N-Cad and CXCL12
What do SAOS2 cells express?
N-Cadherin
What 4 problems plagued the adhesion assay?
- Fixing kills cells, dead cells detach
- Triton X poorly homogenised
- Single laser point of read
- Adhesion substrate cells were large and took up a lot of Crystal Violet
Giving AMD3100 increases circulating MC number in vivo
and enhances action of Bortezomib
A suitable cell type for further E-Selectin experiments?
HUVECS
Name of E-Selectin further work appliance?
Re-circulation parallel flow plate (or microfluidic chip)
What induces sialytion of CD162?
Hypoxia
