Tina:MESC brainbow line: Difference between revisions
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>TinaLo mNo edit summary |
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| 17 ||P7||6/?/2012 | | 17 ||P7||6/?/2012 | ||
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| 18~ | | 18~19 ||P8||5/31/2012 | ||
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| | | 20~21 ||P8||6/28/2012 | ||
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| 22~28 ||P12||7/31/2012 | |||
|- | |- | ||
| 70~72 ||MEF_batch#2_for nine 35mm dishes||7/5/2012 | | 70~72 ||MEF_batch#2_for nine 35mm dishes||7/5/2012 | ||
Line 34: | Line 36: | ||
**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 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. | ** 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 | **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''' | *'''Batch #2''' | ||
**Date: 7/24/2012 ~ | **Date: 7/24/2012 ~ 8/3/2012 | ||
**Cells: CF-1 MEFs from Global Stem (Cat# GSC-6001G, Lot# 94300639) | **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. | ** 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== | == mES brainbow cells expansion detail== |
Latest revision as of 01:09, 4 August 2012
Cell information[edit]
- Description:
- Growth medium:mouse ES cell culture medium
- Cryopreservation medium:mouse ES cell freezing medium
- Cell storage:
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]
<calendar> name=Tina:LabNotes format=%name/%year-%month-%day date=2012/03/01 view=oneyear </calendar>