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.
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