Culture of 16HBE14o- Cells

2020 ◽  
Author(s):  
Nicole A. McNabb ◽  
Shaun D. McCullough
Keyword(s):  
Environmental Protection Agency ◽  
Bronchial Epithelial Cell ◽  
Epithelial Cell Line ◽  
Cell Passage ◽  
Protection Agency ◽  
Bronchial Epithelial ◽  
Plating Density ◽  
The Us ◽  
Trade Names ◽  
Cell Plating

Abstract This protocol describes the thawing, culturing, and cryopreservation of the human bronchial epithelial cell line 16HBE14o- (referred to as “16HBE”). The attached methods document is a formal version of the information included here. The attached worksheet is a fillable PDF that can be used to maintain cell passage records using this protocol.Please note: Deviation from the three-day passage cycle and cell plating density described here typically results in greater culture and experimental variability.Disclaimer: The contents of this article have been reviewed by the US Environmental Protection Agency and approved for publication and do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use.

Culture of IMR90 Cells

2020 ◽  
Author(s):  
Shaun D. McCullough
Keyword(s):  
Environmental Protection Agency ◽  
Human Lung ◽  
Fibroblast Cell ◽  
Protection Agency ◽  
Human Lung Fibroblast ◽  
Plating Density ◽  
Lung Fibroblast Cell Line ◽  
The Us ◽  
Trade Names ◽  
Cell Plating

Abstract This protocol describes the thawing, culturing, and cryopreservation of the human lung fibroblast cell line IMR90. The attached methods document is a formal version of the information included here. The attached worksheet is a fillable PDF that can be used to maintain cell passage records using this protocol. Please note: Deviation from the three-day passage cycle and cell plating density described here typically results in greater culture and experimental variability.Disclaimer: The contents of this article have been reviewed by the US Environmental Protection Agency and approved for publication and do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use.

scholarly journals Establishing Differentiated Air-Liquid Interface Primary Human Bronchial Epithelial Cell Cultures

2021 ◽  
Author(s):  
Lisa Dailey ◽  
Shaun D. McCullough
Keyword(s):  
Epithelial Cell ◽  
Environmental Protection Agency ◽  
Bronchial Epithelial Cell ◽  
Human Bronchial Epithelial Cell ◽  
Protection Agency ◽  
Bronchial Epithelial ◽  
The Us ◽  
Trade Names ◽  
Air Liquid Interface ◽  
Primary Bronchial Epithelial Cells

Abstract This protocol describes the establishment and maintenance of differentiated primary human bronchial epithelial cell (pHBEC) cultures from primary bronchial epithelial cells using Lonza and Gibco media. Note: This is a historical protocol. At the time of publication, this protocol has been superseded by a different version in the McCullough lab; however, it is being published to support the transparency and reproducibility of other studies by which it is referenced.Disclaimer: The information presented here has been reviewed and approved for publication by the US Environmental Protection Agency do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use.

Trans-Epithelial Exposure Model (TEEM) - Epithelial Cells and Fibroblasts

2020 ◽  
Author(s):  
Shaun D. McCullough
Keyword(s):  
Particulate Matter ◽  
Epithelial Cells ◽  
Cell Line ◽  
Environmental Protection Agency ◽  
Airway Epithelium ◽  
Exposure Model ◽  
Protection Agency ◽  
The Us ◽  
Trade Names

Abstract This exposure model is meant to serve as an in vitro representation of trans-epithelial effects of airway diesel and particulate matter exposures on fibroblasts in the stroma. The epithelial cells in the airway epithelium are represented by 16HBE cells and the fibroblasts are represented by either the IMR90 cell line or primary fibroblasts.Disclaimer: The contents of this article have been reviewed by the US Environmental Protection Agency and approved for publication and do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use.

LIVE/DEAD™ Fixable Near-IR Cell Viability Assay

2020 ◽  
Author(s):  
Samantha C. Faber ◽  
Shaun McCullough
Keyword(s):  
Cell Viability ◽  
Environmental Protection Agency ◽  
Protection Agency ◽  
Cell Viability Assay ◽  
Antibody Staining ◽  
Viability Assay ◽  
Near Ir ◽  
The Us ◽  
Trade Names ◽  
Formal Version

Abstract This protocol is used to determine the viability of cells prior to the fixation and permeabilization required for intracellular antibody staining or prior to elimination of biohazardous materials using formaldehyde fixation. The attached methods document is a formal version of the information included here.Disclaimer: The contents of this article have been reviewed by the US Environmental Protection Agency and approved for publication and do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use.

Measurement of Trans-Epithelial Electrical Resistance with EVOM2 and Chopstick Electrode

2020 ◽  
Author(s):  
Shaun D. McCullough
Keyword(s):  
Environmental Protection ◽  
Environmental Protection Agency ◽  
Electrical Resistance ◽  
Cell Monolayer ◽  
Protection Agency ◽  
Commercial Products ◽  
The Us ◽  
Trade Names ◽  
Agency Policy

Abstract Trans-epithelial Electrical Resistance (TEER) can be used as a measure of cell monolayer confluence, health, and integrity. An Epithelial Voltohmmeter (EVOM) is used to take TEER measurements. This method details how to take TEER readings using an EVOM with chopstick electrode.Please note that chopstick electrodes are only designed for use with 12 mm inserts.Disclaimer: The contents of this article have been reviewed by the US Environmental Protection Agency and approved for publication and do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use.

scholarly journals Culture of Primary Human Tracheobronchial Epithelial Cells

2021 ◽  
Author(s):  
Lisa Dailey ◽  
Shaun D. McCullough
Keyword(s):  
Epithelial Cells ◽  
Environmental Protection ◽  
Environmental Protection Agency ◽  
Protection Agency ◽  
Commercial Products ◽  
Brush Biopsy ◽  
The Us ◽  
Trade Names ◽  
Agency Policy

Abstract This protocol is intended for culture of primary human tracheobronchial epithelial cells (pHBEC) obtained by brush biopsy during clinical bronchoscopy, or purchased commercially, using Lonza-based medium.Note: This is a historical protocol. At the time of publication, this protocol has been superseded by a different version in the McCullough lab; however, it is being published to support the transparency and reproducibility of other studies by which it is referenced.Disclaimer: The information presented here has been reviewed and approved for publication by the US Environmental Protection Agency do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use.

scholarly journals Culture and Differentiation of Primary Human Tracheobronchial Epithelial Cells Using STEMCELL Technologies Pneumacult Media

2021 ◽  
Author(s):  
Lisa A. Dailey ◽  
Shaun D. McCullough
Keyword(s):  
Epithelial Cells ◽  
Environmental Protection ◽  
Environmental Protection Agency ◽  
Protection Agency ◽  
Commercial Products ◽  
Brush Biopsy ◽  
Previous Version ◽  
The Us ◽  
Scope Of Application ◽  
Trade Names

Abstract NOTE: A PDF methods document is attached in the supplementary materials.SCOPE OF APPLICATION (LIMITATIONS)This method describes the culture and differentiation of primary human tracheobronchial epithelial cells (pHBEC). Cells used for this method can be obtained by brush biopsy during clinical bronchoscopy or purchased commercially. This method replaces the previous version described in (Dailey and McCullough, 2021a; b). Disclaimer: The contents of this article have been reviewed by the US Environmental Protection Agency and approved for publication and do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use.

scholarly journals Culture of IMR90 Cells in Advanced MEM with Reduced Serum

2021 ◽  
Author(s):  
Nicholas M. Mallek ◽  
Shaun D. McCullough
Keyword(s):  
Environmental Protection Agency ◽  
Human Lung ◽  
Fibroblast Cell ◽  
Lung Fibroblast ◽  
Protection Agency ◽  
Human Lung Fibroblast ◽  
Lung Fibroblast Cell Line ◽  
The Us ◽  
Scope Of Application ◽  
Trade Names

Abstract NOTE: Methods document and worksheet versions of this method are attached as PDFs.SCOPE OF APPLICATION (LIMITATIONS)This method describes the thawing, culturing, and cryopreservation of the human lung fibroblast cell line IMR90 in Advanced MEM-based growth medium. Disclaimer: The contents of this article have been reviewed by the US Environmental Protection Agency and approved for publication and do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use.

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