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Table 2 Characteristics and potentials of three airway epithelial culture methods

From: Patient-specific three-dimensional explant spheroids derived from human nasal airway epithelium: a simple methodological approach for ex vivo studies of primary ciliary dyskinesia

  Crude immediate brush material [21] 2-step ALI [5, 8, 22, 23] 2-step suspension [2, 3, 24, 25] 3D-E spheroids [9, 13]
Characteristics
 Sampling of cells Nasal brush biopsy Nasal brush biopsy Removal of nasal polyp or nasal forceps biopsy Removal of nasal polyp/nasal brush biopsy
 Anesthesia required No No Yes Yes (polyp removal)
No (nasal brush)
 Preparation steps Cell collection
Cell dispersion
Cell collection
Cell dispersion
Cell proliferation Ciliary regeneration
Tissue washing
Enzymatic tissue digestion
Cell dispersion
Cell proliferation
Ciliary regeneration
Cell collection
Cell dispersion
Spheroid formation
 Longevity of cells/culture 1–3 days >1 month >1 month At least 16 days
 Time from sampling to analysis Immediately 4 weeks 4–6 weeks 24 h
 Materials and reagents Falcon tube
NaCl
Glass slide
Falcon tube
Culture medium
T25/75 flask
24-well plate
Transwell insert filters
D-PBS
Trypsin–EDTA
Trypsin-inhibitor
Centrifuge
CO2 incubator
Falcon tube
Pronase
Culture medium
Plastic plate
Collagen gel
Collagenase
T25/75 flask
Gyrotori shaker
CO2 incubator
Falcon tube
Culture medium
24-well plate
CO2 incubator
 Complexity Simple Complex Complex Simple
 Terminally differentiated cells Yes No No Yes
 Cell divisions No Yes Yes No
 Ciliogenesis No Yes Yes No
Known applications
 Diagnostics Yes Yes Yes Yes
 Study of transport mechanisms No Yes No Yes
 Study of ciliary physiology Yes Yes Yes Yes
Potential applications
 Study of drug effects No Yes Yes Yes
  1. Ciliogenesis from suspension, ciliogenesis from air–liquid interface (ALI) and 3D-E spheroids