PneumaCult™-ALI-S Medium

Serum- and BPE-free medium for human small airway epithelial cells cultured at the air-liquid interface
PneumaCult™-ALI-S Medium

Serum- and BPE-free medium for human small airway epithelial cells cultured at the air-liquid interface

1 Kit
Catalog # 05050
Required Products
  1. PneumaCult™-Ex Plus Medium
    PneumaCult™-Ex Plus Medium

    Serum- and BPE-free medium for expansion of primary human airway epithelial cells

  2. Hydrocortisone Stock Solution|07925
    Hydrocortisone Stock Solution

    Cell culture supplement

  3. Animal Component-Free Cell Dissociation Kit
    Animal Component-Free Cell Dissociation Kit

    Dissociation kit for human stem and progenitor cells

  4. Heparin Solution
    Heparin Solution

    Cell culture supplement

Overview

PneumaCult™-ALI-S Medium (Catalog #05050) is a serum- and BPE-free medium for the culture of human small airway epithelial cells at the air-liquid interface (ALI). Small airway epithelial cells cultured in PneumaCult™-ALI-S Medium undergo extensive mucociliary differentiation to form a cuboidal epithelium that exhibits morphological and functional characteristics similar to those of the human small airway in vivo.

Together, PneumaCult™-ALI-S Medium and PneumaCult™-Ex Plus Medium (#05040) constitute a fully integrated BPE-free culture system for in vitro human small airway modeling. This robust and defined system is a valuable tool for basic respiratory research, toxicity studies, and drug development.
Advantages
• Human small airway epithelial cells (HSAEC) cultured in PneumaCult™-ALI-S undergo extensive mucociliary differentiation to form a cuboidal epithelium that closely resembles the small airway epithelium
• PneumaCult™-ALI-S is serum-free and BPE-free to minimize variability
• PneumaCult™-ALI-S and PneumaCult™-Ex Plus constitute a complete, optimized system for expansion, maintenance, and differentiation of HSAEC
Components
  • PneumaCult™-ALI-S Basal Medium, 450 mL
  • PneumaCult™-ALI-S Supplement (10X), 50 mL
  • PneumaCult™-ALI-S Maintenance Supplement (100X), 5 x 1 mL
Subtype
Specialized Media
Cell Type
Airway Cells
Species
Human
Application
Cell Culture, Differentiation, Maintenance
Brand
PneumaCult
Area of Interest
Disease Modeling, Drug Discovery and Toxicity Testing, Epithelial Cell Biology, Respiratory Research
Formulation
Serum-Free

Related Products

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
PneumaCult™-ALI-S Medium
Catalog #
05050
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
PneumaCult™-ALI-S Medium
Catalog #
05050
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
PneumaCult™-ALI-S Medium
Catalog #
05050
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
PneumaCult™-ALI-S Medium
Catalog #
05050
Lot #
All
Language
English

Educational Materials(17)

Brochure
PneumaCult™-ALI-S Medium: Model the Regional Specificity of the Small Airway
Brochure
PneumaCult™-Ex Plus: Generate More Airway Epithelial Cells for Extended Passages
Brochure
Modeling Cystic Fibrosis Airway: PneumaCult™-Ex Plus and PneumaCult™-ALI
Technical Bulletin
Air-Liquid Interface Culture for Respiratory Research
Special Protocol
How to Perform Immunocytochemistry (ICC) Staining of Epithelial Cells Cultured as Monolayers or at the Air-Liquid Interface
Video
2:13
How to Model the Human Airway at the Air-Liquid Interface: Introduction
Video
3:02
Correlating TEER Values with Air-Liquid Interface Culture Morphology
Video
7:42
How to Model the Human Airway at the Air-Liquid Interface: Expansion of HBECs
Video
2:01
Air-Liquid Interface (ALI) Culture of Bronchial Epithelial Cells with PneumaCult™-ALI Medium
Video
5:03
How to Model the Human Airway at the Air-Liquid Interface: Differentiation of HBECs
Video
3:55
Why Use Air-Liquid Interface Cultures for Respiratory Research: Q&A with Dr. Wadsworth
Video
4:25
How to Perform a TEER Measurement to Evaluate Epithelial Barrier Integrity in ALI Cultures
Webinar
12:13
Optimization of Normal Human Bronchial Epithelial Cell 3D Cultures for In Vitro Lung Model Studies
Webinar
18:41
In Vitro Human Airway Modeling Using Primary Airway Epithelial Cells
Webinar
6:56
Studying Respiratory Viruses with ALI Cultures
Webinar
51:30
Studying Cystic Fibrosis Using Primary Human Nasal Epithelial Cells
Mini Review
Airway Epithelial Cells of the Human Respiratory System
Load More Educational Materials

Product Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Research Area
Workflow Stages

Data and Publications

Data

PneumaCult™ Culture System Workflow for Small Airway Research

Figure 1. Overview of the PneumaCult™ Culture System for Small Airway Research

Expansion of human small airway epithelial cells (HSAEC) in submerged culture is performed with PneumaCult™-Ex Plus Medium. During the early Expansion Phase of the air-liquid interface (ALI) culture procedure, PneumaCult™-Ex Plus 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 PneumaCult™-ALI-S is added to the basal chamber only. Differentiation into a mucociliary epithelium is obtained following 21+ days of incubation and can be maintained for more than one year.

Higher proliferation rate of HSAEC cultured in PneumaCult™-Ex Plus Medium compared with other.

Figure 2. HSAEC and HBEC Grow at a Higher Rate During Expansion When Cultured in PneumaCult™-Ex Plus Medium

Human small airway epithelial cells (HSAEC) and human bronchial epithelial cells (HBEC) cultured in PneumaCult™-Ex Plus Medium exhibited higher proliferation rate at every passage compared with cells cultured in Small Airway Epithelial Cell Growth Medium. Cryopreserved HSAEC were obtained commercially at passage 2 (P2) while HBEC were obtained at P1.

HSAEC cultured in PneumaCult™-ALI-S differentiate to form a thin, cuboidal epithelium representative of the small airway.

Figure 3. HSAEC Cultured at the ALI Using PneumaCult™-ALI-S Medium Differentiate to Form a Morphology Representative of the Small Airway Epithelium

Hematoxylin and eosin (H&E) staining of HSAEC and HBEC cultured in PneumaCult™-ALI-S or PneumaCult™-ALI Medium at P3, after 28 days. HSAEC differentiated at the ALI in PneumaCult™-ALI-S formed a thin, cuboidal epithelial layer representative of the in vivo small airway epithelium while HBEC differentiated in PneumaCult™-ALI formed a pseudostratified epithelium resembling the in vivo bronchial epithelium. The ALI cultures were fixed, paraffin-embedded, sectioned, and stained with H&E. All images were taken using a 40X objective. Insert membrane was 10 μm in thickness. Scale bar = 20 μm.

Small airway epithelium markers, SCGB1A1, SCGB3A2, were detected in HSAEC cultured in PneumaCult™-ALI-S Medium.

Figure 4. Small Airway Epithelium Markers Were Detected in HSAEC Cultured in PneumaCult™-ALI-S Medium

Confocal images of whole mount immunostained ALI cultures showing HSAEC and HBEC cultured in PneumaCult™-ALI-S or PneumaCult™-ALI Medium at P3, after 28 days. The ALI cultures were fixed and stained with antibodies for ciliated cells (AC-tubulin; green), club cells (SCGB1A1; magenta), and secretory protein (SCGB3A2; red). The nuclei were counterstained with DAPI (blue). Small airway epithelium markers, SCGB1A1 and SCGB3A2, were detected at higher levels in HSAEC cultured in PneumaCult™-ALI-S compared with HSAEC cultured in PneumaCult™-ALI and HBEC cultured in either PneumaCult™-ALI-S or PneumaCult™-ALI. All images were taken using a 63X objective.

Relative expression of SCGB1A1 and SCGB3A2 was higher in HSAEC cultured in PneumaCult™-ALI-S compared to PneumaCult™-ALI.

Figure 5. Relative Expression of Small Airway Epithelium Markers by qPCR Were Detected at Higher Levels in HSAEC Cultured in PneumaCult™-ALI-S Medium Compared with HSAEC Cultured in PneumaCult™-ALI

HSAEC and HBEC cultured in PneumaCult™-ALI-S or PneumaCult™-ALI Medium at P3. After 28-days of differentiation, the ALI cultures were analysed for small airway epithelium markers, SCGB1A1 and SCGB3A2. Gene of Interest expression was normalized to housekeeping gene, TBP, and expressed as relative quantity (RQ). Relative expression of SCGB1A1 and SCGB3A2 was higher in HSAEC cultured in PneumaCult™-ALI-S Medium compared with HSAEC cultured in PneumaCult™-ALI and HBEC cultured in either PneumaCult™-ALI-S or PneumaCult™-ALI. Relative expression of SCGB3A2 was not detectable in HBEC cultured in either PneumaCult™-ALI or PneumaCult™-ALI-S.

Publications (1)

Scientific reports 2020 Development of a miniaturized 96-Transwell air-liquid interface human small airway epithelial model. T. Bluhmki et al.

Abstract

In order to overcome the challenges associated with a limited number of airway epithelial cells that can be obtained from clinical sampling and their restrained capacity to divide ex vivo, miniaturization of respiratory drug discovery assays is of pivotal importance. Thus, a 96-well microplate system was developed where primary human small airway epithelial (hSAE) cells were cultured at an air-liquid interface (ALI). After four weeks of ALI culture, a pseudostratified epithelium containing basal, club, goblet and ciliated cells was produced. The 96-well ALI cultures displayed a cellular composition, ciliary beating frequency, and intercellular tight junctions similar to 24-well conditions. A novel custom-made device for 96-parallelized transepithelial electric resistance (TEER) measurements, together with dextran permeability measurements, confirmed that the 96-well culture developed a tight barrier function during ALI differentiation. 96-well hSAE cultures were responsive to transforming growth factor $\beta$1 (TGF-$\beta$1) and tumor necrosis factor $\alpha$ (TNF-$\alpha$) in a concentration dependent manner. Thus, the miniaturized cellular model system enables the recapitulation of a physiologically responsive, differentiated small airway epithelium, and a robotic integration provides a medium throughput approach towards pharmaceutical drug discovery, for instance, in respect of fibrotic distal airway/lung diseases.
View All Publications

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