Dear Authors:
I am keen to follow your protocol for generating megakaryocytic progenitors from hiPSCs. My question is with regards to the following text:
"After 10 days, 72 EBs were transferred to each well of a 6 well flat-bottom tissue culture plate containing 7.2 ml of BPEL supplemented with 20 ng/ml rh thrombopoietin (TPO), 25 ng/ml rhSCF and 25 ng/ml rh interleukin (IL)-3 (Peprotech)."
72 EBs in 7.2 ml of medium sounds too much for ONE WELL of a six-well plate! Can you please confirm that this is true? In the Ng ES Nature Protocols paper which you have followed, they transfer 30 EBs in 3 ml of APEL medium per well of six-well plate on day 10.
I would be grateful if you can kindly reply to either sayandip@gmail.com or sayandip.mukherjee@ucl.ac.uk.
Thanks and Regards,
Sayandip Mukherjee.
UCL, London, UK.