Cell line engraftment

[Aparajitha Vaidyanathan]

Hi everyone,

It says in the OG protocol that the BM organoids remain viable for over 45 days. What media conditions are recommended after engraftment with any leukemia cell lines. Will the organoids remain healthy and viable in a leukemia specific media, or it is advisable to use the the organoid specific Phase IV media?

Really appreciate this community. 

Kind regards, 

Aparajitha

[Abdullah Khan]

HI Aparjitha, 

We've tried a few different cocktails and get different levels of survival. What leukaemia specific media were you thinking of? 

My general impression is you can be flexible with your media provided: 

- Avoid DMEM/F12 as base (sugars too high, not enough trace material, preference is for IMDM). 

- There is a reasonably amount of Insulin-transferrin 

- You can strip growth factors if you like, but always keep at least 1ng/TPO. You will get more differentiation if you supplement with some of the essentials (e.g. 5ng/mL SCF, FLT3L). It depends a bit on your growth time line and your readouts. 

Otherwise you can stick to Phase IV media (StemPro-34 base) - that has a good consistent result. 

Abs

[Aparajitha Vaidyanathan]

Hi Abs, 

Thank you for the detailed information and tips. I am looking to use either IMDM or RPMI, but will try optimizing with Phase IV media as well.

Best, 

Aparajitha