Correction for “Novel immortalized human vocal fold epithelial cell line: In vitro tool for mucosal biology,” by Xia Chen, Vlasta Lungova, Haiyan Zhang, Chitrasen Mohanty, Christina Kendziorski, and Susan L. Thibeault, which was first published on January 11, 2021; 10.1096/fj.202001423R (FASEB J. 35, e21243).
The authors report that there are two errors in the Materials and Methods section of the published article. The composition of epithelium cell culture medium (Section 2.1) and stratified DMEM (Section 2.6) are not correctly reported. These errors were present in the manuscript text submitted for production. The authors apologize for these errors.
The corrected passages are as follows:
2.1 | Cells and culture
Primary normal hVFE (E67 and E80) was isolated from human true VF harvested from 67‐year‐old female and 80‐year‐old male donors approximately 10–24 h from death. The Institutional Review Board of the University of Wisconsin‐Madison approved the collection of tissue. For the culture of hVFE from the tissue explant, cell culture plates were coated with rat tail collagen I 30 μg/ml, fibronectin 10 μg/ml, and BSA 10 μg/ml. After a 1‐h incubation to allow proteins to adsorb to the surface, the remaining solution was removed, and the surface was washed twice with PBS. The epithelial layer was carefully removed from the remainder of the tissue with sterile scissors and forceps; the epithelium was rinsed with 70% ethanol and PBS, cut into smaller pieces and placed on the pre‐coated plates. Epithelial cell culture medium (EPC) was composed of Airway Epithelial Cell Basal Medium (PCS‐300‐030; ATCC, Manassas, VA) supplemented with Bronchial Epithelial Cell Growth Kit (PCS‐300‐040; ATCC), cholera toxin 25 ng/ml (Sigma, St. Louis, MO), and primocin 100 ng/ml (InvivoGen, San Diego, CA). After 10–14 days in primary culture, cells were passaged using low‐concentration trypsin–EDTA (0.05% trypsin and 0.02% EDTA) for removing, contaminated fibroblasts. After two to three passages, hVFE (Figure 1) achieved 70%–90% confluence and was used for downstream assays.
2.6 | Creation of a human VF mucosal construct
Prior to seeding on constructs, immortalized hVFE (passages 10–20) was cultivated in the DMEM‐stratified medium that was composed of DMEM/F12 medium with Glutamax (Gibco, Co Dublin, Ireland), supplemented with 2% B27 and 1% N2 (Gibco), 0.4 μg/ml hydrocortisone (Millipore Sigma), 8.4 ng/ml cholera toxin (Sigma), 5 μg/ml insulin (Millipore Sigma), 24 μg/ml adenine (Millipore Sigma), 20 ng/ml EGF, 100 U/ml penicillin, and 0.01 mg/ml streptomycin sulfate (Invitrogen, Carlsbad, CA).