EB/aggregate formation

[Abdullah Khan]

Hi all, 

I've mentioned this to a few of you in follow up zooms/emails - apologies for the delay in uploading it. 

I've never been super happy with the EB generation, so we've been experimenting with improving this. 

Attached is an adapted Aggrewell (Stem Cell Tech) protocol which we've validated and works quite well. 

You use the Aggrewell plates to make a set number of aggregates of specific cell number and then collect these as per usual for day -1. The benefit is the aggregates are evenly sized and you go in knowing exactly how many you have. 

I'd suggest doing a quick 'test' run to validate the approach produces your cell types of interest, but from what we've seen the protocol has identical yields and is just more consistent. 

I suspect there will be some tweaking to do with input cell size for some lines, but so far we've used this on SCT, Gibco, BU repository lines, KOLFJ2.1 and MCNDs with good results! 

Have a good Christmas and New Year from all of us here at TOOT, 

Abs