Tina:MESC brainbow line

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Revision as of 01:09, 4 August 2012 by >TinaLo
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Cell information[edit]

All mES brainbow cells are stored in the N2(l) tank at CERC. (Rack #2, Box position: 8th shelf, Name: ZhangLab Box 3)

Location Passage Date
1~8 P5 5/21/2012
9~16 P6 5/24/2012
17 P7 6/?/2012
18~19 P8 5/31/2012
20~21 P8 6/28/2012
22~28 P12 7/31/2012
70~72 MEF_batch#2_for nine 35mm dishes 7/5/2012
74~81 MEF_batch#1_for four 35mm dishes 6/28/2012

MEF conditioned medium preparation[edit]

  • Purpose: To prepare MEF conditioned medium for feeder free culture system
  • Batch #1
    • Date: 5/29/2012 ~ 6/13/2012
    • Cells: MEFs from Dr. Yang Xu’s Lab; CF-1 MEFs from Global Stem (Cat# GSC-6001M, Lot# 94300663)
    • Exp started on 5/29. MEF cells seed on 5/25 were used. Cell density: 2.5*10^4 cells/cm2. 10 wells. Collect MEF conditioned medium every 24 hrs (106 mL in total) from 5/30 to 6/4. Store at -20゚C.
    • Another exp started on 5/31. Thaw one vial of MEF (from Dr. Xu's Lab), seed in one T75 flask. Cell density: 4*10^4 cells/cm2. (Anticipated production: theoretically 0.3-0.4 mL/cm2). Collect MEF conditioned medium every 24 hrs (106 mL in total) from 6/2 to 6/8. Store at -20゚C.
    • Another exp started on 6/4. Thaw one vial of MEF (CF-1 MITC treated, from GlobalStem) in one T75 and 2 wells of one 6-well plate. Cell density :~4*10^4 cells/cm2. Collect MEF conditioned medium every 24 hrs (106 mL in total) from 6/7 to 6/13. Store at -20゚C.
    • Pool all of the MEF conditioned medium and filter through a 0.22 um filter. Aliquot (40mL/tube) and store at -20゚C. Conditioned medium are good for one month.
  • Batch #2
    • Date: 7/24/2012 ~ 8/3/2012
    • Cells: CF-1 MEFs from Global Stem (Cat# GSC-6001G, Lot# 94300639)
    • Exp started on 7/24. Thaw 1 vial of CF-1 MEFs on to twenty 35mm-wells. Cell density: 2.5*10^4 cells/cm2. Collect MEF conditioned medium every 24 hrs (2*14=28 mL in total/day) from 7/25 to 8/3. Store at -20゚C.
    • Pool all of the MEF conditioned medium and filter through a 0.22 um filter. Aliquot (40mL/tube) and store at -20゚C. Conditioned medium are good for one month.

mES brainbow cells expansion detail[edit]

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