Media for primary cells (Lonza-based)

Media for primary bronchial and sinonasal cells:
  • Base growth media for submerged cells (Human Growth media; HG):
  • 50% Lonza bronchial epithelial basal media (BEBM; CC-3171)
  • 50% MEME (Invitrogen 11905072; Cell center 1154)
  • Supplemented 1x Pen/Strep, 2% Corning NuSerum
  • Add BEGM singlequot components (1 kit per 500 ml total volume):
    • Bovine Pituitary Extract (BPE)
    • Hydrocortisone
    • hEGF
    • Epinepherine
    • Transferrin
    • Insulin
    • Retinoic Acid
    • DO NOT ADD Triiodothyronine (T3) or Gentamycin-Amphotericin B

Aliquot media into 50ml brown tubes. 
Do not warm more media than necessary. 
Store at 4°C in the dark. 

For ciliation, supplement media with B-ALI inducer (retinoic acid).  Add inducer fresh each time you feed to only as much media as you need at one time.  
Aliquot B-ALI inducer (SingleQuots No. 00556823; contains 1.0 ml stock solution) into 25 uL aliquots and freeze in the dark at -20.
 Add 25 uL inducer to 13 uL HG to make (Human Differentiation media; HD).