Purecol bovine collagen solution

All primary HPCa samples were obtained with written informed patient consent in accordance with federal and institutional guidelines, which was approved by M.D. Anderson Cancer Center Institutional Review Board (IRB) (Protocol number LAB04-0498) and the Roswell Park Comprehensive Cancer Center IRB (BDR073416). Three HPCa samples were utilized in this study: HPCa229 (71 years old, Gleason grade 3 + 4 = 7, 30% tumor involvement), HPCa231 (72 years old, Gleason grade 4 + 4 = 8, 100% tumor involvement), and HPCa232 (72 years old, Gleason grade 4 + 3 = 7, 100% tumor involvement). Basic procedures for patient HPCa sample processing have been described previously^{11 (link),14 (link)}. Single HPCa epithelial cells were purified into Trop2⁺CD45⁻ cell population using flow cytometry (FACS), cultured in PureCol bovine collagen solution (Advanced BioMatrix) coated dishes^{16 (link)} in modified WIT medium^{25 (link)}, and treated with DMSO (vehicle control) or Enza (10 or 20 μM).

Human bronchial epithelial (HBE) cells from 6 donors were obtained from Dr. Scott Randell (University of North Carolina, Chapel Hill, NC, USA) and cultured as described previously [56 (link)]. Briefly, cells were grown in BronchiaLife basal medium (Lifeline Cell Technology, Frederick, MD, USA) supplemented with the BronchiaLife B/T LifeFactors Kit (Lifeline) as well as 10,000 U/ml Penicillin and 10,000 μg/ml Streptomycin. Culture plates were coated with PureCol Bovine Collagen Solution (Advanced Bio Matrix, Carlsbad, CA, USA). For OMV exposures, HBE cells were polarized on PureCol-coated permeable supports (#3801 Corning Inc., Corning, NY, USA) at an air-liquid interface for 3–4 weeks. During polarization, HBE cells were supplemented on the basolateral side with Air Liquid Interface (ALI) medium [57 (link)].