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Tina:Cardiomyocyte differentiation in H9 ESCs (CDH9)
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==Experiment #2 (CDH9_E1_T2)== *Study Purpose: Try 2 different induction medium and 2 different cell density during differentiation and see how it works on H9 ESCs directed cardiomyocyte differentiation. *Date: 4/20/2013 ~ 5/20/2013 *Cells: H9 ESC line, Brainbow Clone #3 (H9Bb3). Cells seeded on 4/20. ('''1:3''' dilution from the second passage of H9 ESCs under a feeder free culture system). *Protocol used: Christian Metallo Lab protocol. *Two induction medium used: Gln+ induction medium,DMK induction medium *Two cell density used: 100%, 70% confluency. '''24-well plate #1''' {| {{table}} border=1 | align="center" style="background:#f0f0f0;"| | align="center" style="background:#f0f0f0;"|'''1''' | align="center" style="background:#f0f0f0;"|'''2''' | align="center" style="background:#f0f0f0;"|'''3''' | align="center" style="background:#f0f0f0;"|'''4''' |- | align="center" style="background:#f0f0f0;"|A||100%, Gln, repeat 1||100%, Gln, repeat 2||100%, DMK, repeat 1||100%, DMK, repeat 2 |- | align="center" style="background:#f0f0f0;"|B||100%, Gln, repeat 3||100%, Gln, repeat 4||100%, DMK, repeat 3||100%, DMK, repeat 4 |- | align="center" style="background:#f0f0f0;"|A||70%, Gln, repeat 1||70%, Gln, repeat 2||70%, DMK, repeat 1||70%, DMK, repeat 2 |- | align="center" style="background:#f0f0f0;"|B||70%, Gln, repeat 3||70%, Gln, repeat 4||70%, DMK, repeat 3||70%, DMK, repeat 4 |- |} *Induction: 4/23 (Day 0). Cells reached 100% and 70% confluency, respectively. Start experiment. Mix CHIR99021 and DMK with medium before adding to each well. *4/24 (Day 1). Change medium (pre-mix DMK). Still, observed lots of cell round-up. May be died or detached. can't tell. *4/25~5/20. Change medium accordingly. Take pictures. *Pictures: *Perform flow analysis on 5/20. Results: Cell numbers were too low. The test was not successful. Flow analysis will be repeat next time.
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