Song Chen:Daily/NOTES/2014-3-6

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hES change medium:

  1. PGP iPS Brainbow knockin #1, #2-4 and #2-6 p2, 2 well*6 well plate in hES/NHSM, respectively;
  2. H9 p62 2*6 cm+24 well*48-well plate in NHSM
  3. H9 Bb6 2 well*6 well plate
  4. PGPs iPS LCB+CRE p62 1*6 cm in hES medium


Send H9 in NHSM:

1*T25 flask to UCLA via Fedex(800555872882)


DE differentition(H9 p62, for methylation analysis-Dinh):

3 well*6-well plate, hES at d2, ActA100 d4


check default GFP expression in knockin iPS lines: Setting: Expose 1209 ms, Cerc, 20x obj,GFP/Brightfield, KI(knockin)

KI#1 KI#2-4 KI#2-6 Bb#6(random integrated)
File:030614 KI-1 GFP.png File:030614 KI-2-4 GFP.png File:030614 KI-2-6 GFP.png File:030614 Bb6 GFP.png
File:030614 KI-1 BF.png File:030614 KI-2-4 BF.png File:030614 KI-2-6 BF.png File:030614 Bb6 BF.png
File:030614 KI-1 Merge.png File:030614 KI-2-4 Merge.png File:030614 KI-2-6 Merge.png File:030614 Bb6 Merge.png

Knockin iPS cell lines don't express default GFP in every cells as expected. #2-4 has no detectable GFP expression, while #1 and #2-6 have a few GFP positive cells. And when compared with brainbow random integrated cell line #6, GFP is expressed at a relative low level in knockin cells, which probably due to the nature of single copy insertion of knockin. The possibility of fluctuation in brainbow expression between knockin cells or contamination with non-transgenic iPS cells await further verification.