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Table 2 Summary of maximal TEER values recorded real-time by the CellZscope system for hBMEC mono-cultures using a range of different culture conditions

From: Comparative study of four immortalized human brain capillary endothelial cell lines, hCMEC/D3, hBMEC, TY10, and BB19, and optimization of culture conditions, for an in vitro blood–brain barrier model for drug permeability studies

Culture conditions Cell seeding density (cells/cm2) Time (d) Mean max. TEER ± S.E.M. (Ωcm2)
Cell seeding density: 3.0 × 104 cells/cm2, coating: collagen type I (10 μg/cm2) 3.0 × 104 1.72* 25.9 (± 0.635)
Cell seeding density: 4.5 × 104 cells/cm2, coating: collagen type I (10 μg/cm2) 4.5 × 104 3.19 43.6 (± 3.89)
Growth medium with additional HC (500 nM) for 3 days (only apical), coating: collagen type I (10 μg/cm2) 4.5 × 104 3.19 43.2 (± 4.27)
Growth medium with additional HC (1500 nM) for 3 days (only apical), coating: collagen type I (10 μg/cm2) 4.5 × 104 3.19 40.4 (± 0.646)
Cell seeding density: 6.0 × 104 cells/cm2, coating: collagen type I (10 μg/cm2) 6.0 × 104 1.72* 36.3 (± 2.06)
Cell seeding density: 9.0 × 104 cells/cm2, coating: collagen type I (10 μg/cm2) 9.0 × 104 4.15 38.9 (± 0.928)
Growth medium with dexamethasone (500 nM) for 3 days (only apical), coating: collagen type I (10 μg/cm2) 9.0 × 104 4.15 35.9 (± 0.338)
Growth medium with dexamethasone (1000 nM) for 3 days (only apical), coating: collagen type I (10 μg/cm2) 9.0 × 104 4.15 38.0 (± 1.26)
Growth medium with additional hEGF (50 ng/mL) for 3 days (only apical), coating: collagen type I (10 μg/cm2) 9.0 × 104 4.15 36.4 (± 0.902)
Growth medium with additional hEGF (100 ng/mL) for 3 days (only apical), coating: collagen type I (10 μg/cm2) 9.0 × 104 4.15 40.0 (± 1.94)
Growth medium with 8-(4-CPT)cAMP (250 μM) and RO-20-1724 (17.5 μM) (apical and basolateral), coating: collagen type I (10 μg/cm2) 9.0 × 104 2.84 20.0 (± 0.509)
Growth medium with 8-(4-CPT)cAMP (250 μM) and RO-20-1724 (17.5 μM), added after 2 days in vitro (apical and basolateral), coating: collagen type I (10 μg/cm2) 9.0 × 104 2.84 20.9 (± 1.36)
Cell seeding density: 15 × 104 cells/cm2, coating: collagen type I (10 μg/cm2) 15 × 104 2.75 28.4 (± 2.47)
Coating: collagen type IV/fibronectin (80/20 μg/cm2) 15 × 104 2.75 29.3 (± 1.10)
Coating: matrigel (80 μg/cm2) 15 × 104 2.75 16.6 (± 0.183)
Growth medium with 10% HS (apical and basolateral), coating: collagen type I (10 μg/cm2) 15 × 104 1.26* 20.4 (± 0.867)
Growth medium with 20% HS (apical and basolateral), coating: collagen type I (10 μg/cm2) 15 × 104 1.21* 26.2 (± 0.324)
ACM-1 (apical and basolateral), coating: collagen type I (10 μg/cm2) 15 × 104 1.64* 13.5 (± 0.614)
Mixture of ACM-2 and growth medium 20% FBS (1:1) (apical and basolateral), coating: collagen type I (10 μg/cm2) 15 × 104 3.35 23.8 (± 0.572)
  1. 24-Well tissue culture inserts were from Greiner Bio-one® (transparent PET membrane, pore size 3.0 μm, 0.6 × 106 pores/cm2, n = 2–3). *Before first change of medium.