Cell Culture Guidelines
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Maintenance of Common Cell Lines
Cell Type | Media | Maintenance | Splitting | Notes |
RAW267.4 (Murine macropahge) | DMEM + 10% FBS | Grow to 80% confluency, split 1:5 to 1:10 | Scrape | Prone to genetic drift; use for 20 passages or less |
J774.1 (Murine macropahge) | DMEM + 10% FBS | Grow to 80% confluency, split 1:5 to 1:10 | Scrape | |
THP-1 (Human monocytic) | RPMI 1640 + 10% FBS | Spin to recover, split no ore than 1:15 | Suspension | 0.05 mM 2-mercaptoethanol can be added to enhance growth |
CHO-K1 (Hampster epithelial) | Preferred: Hams F-12 + 10% FBS Alternative: DMEM + 10% FBS |
Grow to 80% confluency, split 1:5 to 1:10 | Trypsin-EDTA | Requires proline in media, therefore RPMI should not be used |
HeLa (Human epithelial) | Preferred: DMEM + 5-10% FBS Alternative: EMEM + 5-10% FBS |
Grow to 80% confluency, split 1:5 to 1:10 | Trypsin-EDTA | |
HEK (Human epithelial) | DMEM + 10% FBS | Grow to 80% confluency, split 1:5 to 1:10 | Trypsin-EDTA | |
12CA5 (Murine hybridoma) | Growth: DMEM or RPMI + 10% HI-FBS Antibody Produciton: Hybridoma SFM |
Grow to high denisty in DMEM; spin to concentrate and resuspend in SFM for antibody production |
Suspension | Supplement growth media with non-essential amino acids, L-glutamine, sodium pyruvate and MEM Vitamins if producing antibody in RMPI or DMEM instead of SFM. |
COS-7 (Monkey epithelial) | DMEM + 10% FBS | Grow to 80% confluency, split 1:5 to 1:10 | Trypsin-EDTA | |
HEK293T (Human epithelial) | DMEM + 10% FBS | Grow to 80% confluency, split 1:5 to 1:10 | Trypsin-EDTA | |
Jurkat (Human T cell) | RPMI + 10% FBS | Grow to 80% confluency, split 1:5 to 1:10 | Trypsin-EDTA | |
C57 (Immortalised peritoneal macrophage from C57/Bl6 mouse) | RPMI + 10% FBS | Grow to 80% confluency, then split 1:5 to 1:10 | Scrape |
Notes:
- FBS = fetal bovine serum (AKA FCS)
- HI-FBS = heat inactivated FBS. Heat-inactivate by heating 50 mL of FBS to 55C for 30 min.
- Antibiotics/mycotics can be added to most cultures, but shoudl be removed for functional assays.