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)
- 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).