BEAS2B cells were plated at a density of 1 x [10.sup.4] cells/well in flat-bottomed 96-well plates containing BEGM
The samples were immediately placed in BEGM
medium (Bronchial Epithelial Cell Growth Media, Clonetics-Lonza) with no serum, supplemented with penicillin 100 U/ml, streptomycin 100 [micro]g/ml, and glutamine.
In contrast to O, significant change of the BEGM
from the distal electrode and ME2 (BEGM-DME2) occurred before and after RFA procedure.
Normal human bronchial epithelial cells (BEAS-2B) were purchased from American Tissue Type Culture Collection (ATCC; CRL-9609) and were cultured in BEGM
media at 37[degrees]C in a humidified incubator with 5% C[O.sub.2].
Abbreviations: ATCC, American type culture collection; BEGM
, Bronchial epithelial cell growth medium; BW, Body weight; Cl, Cell index value; CM2, C.
For MWCNTs, the CPPs performed additional experiments using BEGM
medium (for BEAS-2B cells), showing that the combination of BSA and 1,2-dipalmitoyl-snglycero-3 phosphocholine (DPPC) provided optimal suspension stability (see Supplemental Material, Figure S1A).
The cells were cultured as monolayer in a humidified atmosphere at 37[degrees]C and 5% C[O.sub.2] in flasks, precoated with collagen (collagen IV from human placenta, Sigma-Aldrich) and using bronchial epithelial growth medium (BEGM
by Lonza; singleQuot Kit Lonza) in the presence of 1 [micro]g/ml of puromycin.
was then replaced by Ultroser G medium (USG) (F12/MEM 1: 1, +Supplements Mix +2% USG) to establish an air-liquid interface (ALI) culture system.
Stock solutions of metals were prepared in sterile water (Baxter Healthcare Corp., Deerfield, IL) and were diluted with BEGM
Briefly, bronchial epithelial cells were obtained and seeded in polycarbonate/polyester porous collagen-precoated membranes (PCF Millicell inserts, Millipore, Bedford, MA) using Bronchial Epithelial Cell Growth Medium (BEGM
) (Lonza, Basel, Switzerland) containing 5% fetal bovine serum (FBS) and cultured at 37[degrees]C for approximately 24-48 h.
BEAS-2B cells (1-1.5 x [10.sup.6]) were seeded into T-25 flasks (Coming, Coming, NY) containing bronchial-epithelial growth medium (BEGM
), before expansion in T-150 flasks.
Louis, MO) in bronchial epithelial cell growth medium (BEGM
; Clonetics, San Diego CA) supplemented with bovine pituitary extract, insulin 5 [micro]g/mL, hydrocortisone 0.5 [micro]g/mL, gentamicin 50 [micro]g/mL, retinoic acid 0.1 [micro]g/mL, transferrin 10 [micro]g/mL, triiodothyrodine 6.5 ng/mL, epinephrine 0.5 [micro]g/mL, and human epidermal growth factor 0.5 ng/mL, and dislodged from the brushes by pipetting.