PneumaCult™

PneumaCult™ Culture System for Airway Epithelial Cells

PneumaCult™ is a defined, serum- and bovine pituitary extract (BPE)-free culture system that supports the expansion and differentiation of human airway epithelial cells. Use PneumaCult™-Ex Plus or PneumaCult™-Ex to expand airway epithelial cells in submerged culture. Use PneumaCult™-ALI to achieve extensive mucociliary differentiation at the air-liquid interface (ALI) or in airway organoid culture. Cells expanded and differentiated with the PneumaCult™ system retain key features of the in vivo airway epithelium such as a pseudostratified mucociliated epithelial cell layer consisting of goblet cells, basal cells and motile cilia. ALI cultures provide a physiologically relevant model for studying respiratory biology, infection and disease.
Q&A with Dr. Samuel Wadsworth: Air-Liquid Interface Culture for Respiratory Research

What is Air-Liquid Interface Culture?

Air-liquid interface (ALI) culture is the most physiologically relevant model for studying the respiratory epithelium in vitro. In these cultures, airway epithelial basal cells are seeded in a tissue culture insert with a porous membrane. After cells reach confluence, the medium is removed from the apical chamber to expose the cells to the air. This “air-lift”, in combination with a specialized medium in the basal chamber, triggers basal cell differentiation to produce a pseudostratified mucociliary epithelium that can be used to study cell biology, respiratory disease, toxicity of inhaled substances and more. In this video Dr. Samuel Wadsworth discusses how ALI cultures hold enormous potential for respiratory research.

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Modeling the Human Airway

Primary airway epithelial cells must be cultured at the air-liquid interface in order to recapitulate the pseudostratified mucociliary phenotype observed in vivo. Cells differentiated with the PneumaCult™ culture system exhibit the appropriate three-dimensional architecture with basal cells lining the transwell inserts and ciliated and goblet cells extending to the apical surface. In this video Dr. Michael Riedel provides an overview of PneumaCult™-ALI and how it is optimized for ALI culture.

Why Use PneumaCult™?

  • Primary human airway epithelial cells differentiated with PneumaCult™-ALI closely model the human airway.
  • Defined and BPE-free PneumaCult™ media formulations maximize experimental reproducibility.
  • Simple and optimized protocols make PneumaCult™ media easy to use.

Why Go BPE-Free?

Bovine pituitary extract (BPE) is an undefined component used in several types of serum-free medium. However its presence can lead to experimental variability. Removing BPE reduces the variability and maximizes experimental reproducibility. The PneumaCult™ system has been formulated to be BPE-free while maintaining the highest possible performance of the media. See the results when PneumaCult™-Ex goes head-to-head with a BPE-containing medium.

Why Use PneumaCult™-Ex Plus?

Current feeder-free expansion media for culturing primary human airway epithelial cells can only support a limited number of passages while maintaining robust mucociliary differentiation potential. Unfortunately, this limitation restricts the number of experiments researchers can perform using primary cells. PneumaCult™-Ex Plus is a feeder- and BPE-free culture medium that puts an end to this limitation: researchers can expand cells for a higher number of passages during expansion culture, while maintaining mucociliary differentiation potential during the subsequent ALI culture. Ultimately, PneumaCult™-Ex Plus enables researchers to perform more experiments with a single sample. See the results when PneumaCult™-Ex Plus goes head-to-head with other commercially-available expansion media.

The PneumaCult™ Culture System

PneumaCult™-Ex Plus

PneumaCult™-Ex Plus

Recommended for:

Improved expansion of primary human airway epithelial cells

Species:

Human

Culture Type:

Submerged culture

PneumaCult™-Ex

PneumaCult™-Ex

Recommended for:

Expansion of primary human airway epithelial cells

Species:

Human

Culture Type:

Submerged culture

PneumaCult™-ALI

PneumaCult™-ALI

Recommended for:

Extensive mucociliary differentiation of primary human airway epithelial cells

Species:

Human

Culture Type:

Air-liquid interface (ALI) or sphere culture
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Figure 1. Overview of the PneumaCult™ Culture System

Expansion of human bronchial epithelial cells (HBECs) in submerged culture is performed with PneumaCult™-Ex Plus or PneumaCult™-Ex. During the early “Expansion Phase” of the ALI culture procedure, PneumaCult™-Ex Plus or PneumaCult™-Ex is applied to the apical and basal chambers. Upon reaching confluence, the culture is air-lifted by removing the culture medium from both chambers, and adding PneumaCult™-ALI to the basal chamber only. Differentiation into a pseudostratified mucociliary epithelium is obtained following 21-28 days of incubation and can be maintained for more than one year.

Key Applications

Epithelial Cell Biology

Müller L et al. (2013) Culturing of human nasal epithelial cells at the air liquid interface. J Vis Exp (80): e50646, DOI:10.3791/50646.

Stem Cell Biology

Tata PR et al. (2013) Dedifferentiation of committed epithelial cells into stem cells in vivo. Nature 503(7475): 218-23.

Effects of Pollutants, Cigarette Smoke and Other Toxic Substances

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