Primary Airway Epithelial Cells and Growth Media

These products are available in the US, Canada, and select European countries.

• Primary Airway Epithelial Cell Culture System
• Airway Epithelial Cell Culture System Quality Control
• Materials for Airway Epithelial Cell Culture

Human Primary Airway Epithelial Cell Culture System

The epithelium is the interface between the body and the external environment and covers all exterior surfaces and interior lumina. The diverse functions of epithelial cells include absorption, secretion, protection, transcellular transport, and sensation.

PromoCell^® airway epithelial cells are isolated from normal human adult tissues of the large airways including nasal mucosa, trachea, and bronchi. Shortly after isolation, all human airway epithelial cells are cryopreserved at passage 2 (P2) using a proprietary serum-free freezing medium, Cryo-SFM. After thawing, each cryovial contains at least 500,000 viable cells. Human epithelial cells are also available as proliferating cell cultures made from cryopreserved cells that have been thawed and cultured for three days.

• Cultivation Protocol for Human Airway Epithelial Cells

The Airway Epithelial Cell Growth Medium is a serum-free growth medium optimized for the in vitro cultivation of epithelial cells from large air passages. The Airway Epithelial Cell Growth Medium is optimized for growing primary human cells, but can also be used for equine, porcine, murine, bat and rat airway epithelial cells. This medium is available in two options: Medium (ready-to-use) and Medium Kit. The Medium (ready-to-use) option consists of a 500 mL bottle of Basal Medium and one vial of SupplementMix. The Medium Kit consists of a 500 mL bottle of Basal Medium and the SupplementPack (a set of individual vials with pre-measured supplements), which gives the user full control over the media formulation. Adding the SupplementMix or the SupplementPack to the Basal Medium results in the complete Airway Epithelial Cell Growth Medium. The Basal Medium (with or without phenol red), SupplementMix, and SupplementPack may also be purchased separately.

Airway Epithelial Cell Growth Medium is Recommended for

• Human Nasal Epithelial Cells (HNEpC)
• Human Bronchial Epithelial Cells (HBEpC)
• Human Tracheal Epithelial Cells (HTEpC)

Airway Epithelial Cell Growth Medium Supplementation Details

The Airway Epithelial Cell Growth Medium contains all of the growth factors and supplements needed for the optimal growth of human airway epithelial cells. These include epidermal growth factor, insulin, and transferrin (see the table below). Airway Epithelial Cell Growth Medium does not contain antibiotics or antimycotics and is formulated for use in an incubator with an atmosphere of 5% CO₂

Final Airway Epithelial Cell Growth Medium Supplement Concentrations (After Addition)

Component	Final Concentration
Bovine Pituitary Extract	0.004 mL / mL
Epidermal Growth Factor (recombinant human)	10 ng / mL
Insulin (recombinant human)	5 μg / mL
Hydrocortisone	0.5 μg / mL
Epinephrine	0.5 μg / mL
Triiodo-L-thyronine	6.7 ng / mL
Transferrin (recombinant human)	10 μg / mL
Retinoic Acid	0.1 ng / mL

Preparing Supplemented Media for Primary Airway Epithelial Cells

• Thaw the Airway Epithelial Cell Growth Medium SupplementMix or SupplementPack at 15 – 25 °C.
• Aseptically mix the supplement solutions by carefully pipetting up and down.
• Transfer the entire contents of each supplement to the Airway Epithelial Cell Basal Medium.
• Close the bottle and swirl gently until a homogeneous mixture is achieved.

Storage and Stability of Airway Epithelial Cell Products

• Upon arrival, immediately store the Airway Epithelial Cell Basal Medium at 4 – 8 °C in the dark; do not freeze the medium. Store the Airway Epithelial Cell Growth Medium SupplementMix or SupplementPack at -20 °C.
• The products are stable until the expiration date stated on the label when stored as directed.
• Once supplements are added to the Airway Epithelial Cell Basal Medium, the shelf life of the complete medium is six weeks at 4 – 8 °C. Do not freeze the complete medium.
• For use, prewarm an aliquot of the complete Airway Epithelial Cell Growth Medium and maintain the remaining medium at
  4 – 8 °C.
• After delivery, cryopreserved cells should be stored in liquid nitrogen or seeded directly.
• Proliferating cells must be processed immediately.

Airway Epithelial Cell Culture System Quality Control

Human Primary Airway Epithelial Cells Quality Control

The following rigid quality control tests are performed for each lot of Airway Epithelial Cells:

• Cell morphology, adherence rate, and cell viability testing
• Flow cytometric analyses for cell-type-specific marker cytokeratin
• Growth performance testing through multiple passages, up to 15 population doublings (PD) under culture conditions without antibiotics and antimycotics
• Testing for the absence of HIV-1, HIV-2, HBV, HCV, HTLV-1, HTLV-2 and microbial contaminants (fungi, bacteria, and mycoplasma)
• A detailed certificate of analysis (CoA) is available for each lot.

Airway Epithelial Cell Specifications

Product	Plating Density	Passage after Thawing	Marker	Population Doublings
Human Nasal Epithelial Cells (HNEpC)	10,000 - 15,000 cells per cm²	P2	Cytokeratin+	> 15
Human Tracheal Epithelial Cells (HTEpC)	10,000 - 15,000 cells per cm²	P2	Cytokeratin+	> 15
Human Bronchial Epithelial Cells (HBEpC)	10,000 - 15,000 cells per cm²	P2	Cytokeratin+	> 15

Primary Epithelial Cell Growth Medium Quality Control

All lots of PromoCell^® Airway Epithelial Cell Growth Medium are subjected to comprehensive quality control tests using primary human airway epithelial cells. Each lot is checked for growth-promoting activity, adherence rate, and typical morphology of the tested cells. Approved in-house lots of Airway Epithelial Cell Growth Media are used as a reference. In addition, every lot of media is tested for the absence of microbial contaminants (fungi, bacteria, mycoplasma).

Intended Use of Airway Epithelial Cell Products

These products are for in vitro use only and not for diagnostic or therapeutic procedures. For safety precautions, please see appropriate MSDS.

Note: Due to their low serum content or the absence of serum, PromoCell media are not suitable for trypsin neutralization (when subculturing the cells, for example). Instead, we recommend using our DetachKit (C-41200, C-41210, C-41220), which contains HEPES BSS, Trypsin/EDTA and Trypsin Neutralizing Solution.

Materials for Airway Epithelial Cell Culture