Serum- and BPE-free medium for human small airway epithelial cells cultured at the air-liquid interface
产品优势
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
产品组分包括
PneumaCult™-ALI-S Basal Medium, 450 mL
PneumaCult™-ALI-S Supplement (10X), 50 mL
PneumaCult™-ALI-S Maintenance Supplement (100X), 5 x 1 mL
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.
Area of Interest Disease Modeling, Drug Discovery and Toxicity Testing, Epithelial Cell Biology, Respiratory Research
Formulation Category Serum-Free
Data Figures
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.
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.
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.
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.
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.
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.
Development of a miniaturized 96-Transwell air-liquid interface human small airway epithelial model. T. Bluhmki et al. Scientific reports 2020
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.
Thank you for your interest in IntestiCult™ Organoid Growth Medium (Human). Please provide us with your contact information and your local representative will contact you with a customized quote. Where appropriate, they
can also assist you with a(n):
Estimated delivery time for your area
Product sample or exclusive offer
In-lab demonstration
By submitting this form, you are providing your consent to STEMCELL Technologies Canada Inc. and its subsidiaries and affiliates (“STEMCELL”) to collect and use your information, and send you newsletters and emails in accordance with our
privacy policy. Please contact us with any questions that you may have. You can unsubscribe or change your email preferences at any time.
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.
