PneumaCult™-ALI-S Medium

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

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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
257 USD

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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

Scientific Resources

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 for
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.

PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.