Cell Culture Guidelines

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.