As we continue to passage this sphere (or population of cells) in this manner we would observe a 1-fold expansion, which is represented by a flat growth curve as seen in Figure 1a(i).
Therefore we tested whether additional information is gained from continuing to passage such poor-growing strains.
Another routine that may be successfully employed is to continue to passage the cells in stock flasks kept at 37°C and periodically changed with fresh maintenance medium.
Incubate 1-2 h, shake every 10 min Wash 2 times with 1X PBS Add 300 ÎL PBS Read on the Flow Cytometer * If CD14 is positive, continue to passage cells since macrophages are insensitive to trypsinization.
In early October, the UN Security Council met on the matter of the canal's nationalization and adopted a resolution recognizing Egypt's right to control the canal as long as it continued to allow passage through it for foreign ships.
Once cells had grown to 90-100% confluency they were passaged into a 25-cm2 tissue culture flask and continued to be passaged into larger flasks until sufficient numbers were generated for storage.
Obama and other Democratic leaders, especially Speaker of the House Nancy Pelosi, thought otherwise and continued to push for passage.
She reportedly created makeshift sleeves out of notebook paper, but the House sergeant at arms continued to deny her passage.
Two early hESC lines were obtained, but none of them continued to grow beyond passage two.
After about 2 weeks, when cells were subconfluent, first-passage (P1) cells were detached with trypsin-EDTA (Invitrogen, Paisley, Renfrewshire, UK) and split at a 1 2 ratio; culture was continued to produce second-passage (P2) cells, which were used for experiments.
