How to grow this line
Notes kindly provided by Terri D. Bryson and Aaron Hernandez from Steven Henikoff lab, HHMI/Fred Hutchinson Cancer Research Center.
Using media M3+BPYE+5%FBS we let them grow until there is a fairly confluent monolayer topped by an evenly spaced layer of rounding clumps of cells which easily disperse when pipetted up and down thoroughly. Attached is a picture for reference. We let them grow to about 10-12x10^6cells/mL and split to 3.0x10^6 cells/mL twice a week. This is the 4th round of cells grown in this fashion and they maintain a high viability for at least 11 passages. Aaron learned that if the cells are split any sparser than this the culture will stop growing around passage #7.