PneumaCult™-Ex Medium

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

Interested in trying STEMCELL's products for respiratory research? Fill out this form to request information about introductory offers.

PneumaCult™-Ex Medium

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

From: 206 USD
Catalog #
(Select a product)
Serum- and BPE-free medium for expansion of primary human airway epithelial cells
Add to Wish List

Product Advantages


  • PneumaCult™-Ex is a defined (BPE-free) medium that delivers consistent performance for expansion of HBECs

  • When used together, PneumaCult™-Ex and PneumaCult™-ALI constitute a complete cell culture media system for expansion of primary human airway cells and their subsequent differentiation to a pseudostratified mucociliary epithelium

What's Included

  • PneumaCult™-Ex Basal Medium, 490 mL
  • PneumaCult™-Ex 50X Supplement, 10 mL
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.

Overview

PneumaCult™-Ex is a defined, serum- and BPE-free cell culture medium that supports rapid expansion of human airway epithelial cells.

Primary airway epithelial cells cultured in PneumaCult™-Ex expand rapidly over at least 3 passages while maintaining a cobblestone morphology and uniform expression of the basal cell markers p63 and p75NTR. Additionally, cells cultured in PneumaCult™-Ex can be differentiated to form a pseudostratified mucociliary epithelium when cultured at the air-liquid interface in PneumaCult™-ALI.

Together, PneumaCult™-Ex and PneumaCult™-ALI constitute a fully integrated BPE-free culture system for in vitro human airway modeling. This robust and defined system is a valuable tool for basic respiratory research, toxicity studies, and drug development.

Learn how to culture human airway epithelial cells at the ALI in our On-Demand Pulmonary Course or browse our Frequently Asked Questions (FAQs) about the ALI culture workflow using PneumaCult™.
Subtype
Specialized Media
Cell Type
Airway Cells
Species
Human
Application
Cell Culture, Expansion, Maintenance
Brand
PneumaCult
Area of Interest
Epithelial Cell Biology
Formulation Category
Serum-Free

Data Figures

HBECs Cultured in PneumaCult™-Ex Exhibit Cobblestone Morphology

Figure 1. HBECs Cultured in PneumaCult™-Ex Exhibit Cobblestone Morphology

Commercially available, cryopreserved, passage 1 (P1) HBECs were seeded into PneumaCult™-Ex or a Control medium (BEGM™, Lonza). Cells exhibit cobblestone morphology in both culture media, as seen in representative images of confluent cultures 5 days post-seeding (A,B). HBECs cultured for an additional 3 passages in both PneumaCult™-Ex and Control medium continue to expand and retain their normal cobblestone morphology, as shown by representative images of confluent P4 cultures at 7 days post-seeding (C,D). All images were taken through 10X objective.

HBECs Cultured in PneumaCult™-Ex Exhibit Uniform Expression of Basal Cell Markers

Figure 2. HBECs Cultured in PneumaCult™-Ex Exhibit Uniform Expression of Basal Cell Markers

Passage 3 HBECs cultured in PneumaCult™-Ex demonstrate extensive co-labeling of the basal cell markers p63 (red) and p75NTR (green, A). A representative merged image indicates widespread co-labeling of p63, p75NTR and the nuclear stain DAPI (blue, B).

HBECs Cultured in PneumaCult™-Ex Exhibit Comparable Expansion Rates to Cells Cultured in Control Medium

Figure 3. HBECs Cultured in PneumaCult™-Ex Exhibit Comparable Expansion Rates to Cells Cultured in Control Medium

Commercially available, cryopreserved, P1 HBECs were seeded into PneumaCult™-Ex or a Control medium (BEGM™, Lonza). In seven independent donor samples, the average fold expansion over four passages was not significantly different between cells cultured in PneumaCult™-Ex and cells cultured in the Control medium (7.1 ± 1.4 vs. 7.2 ± 1.9, mean ± SD, n = 7, p = 0.9 in paired t-test).

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Catalog #
05008
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
05008
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
05008
Lot #
All
Language
English

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.

Resources and Publications

Publications (14)

Impact of KLF4 on Cell Proliferation and Epithelial Differentiation in the Context of Cystic Fibrosis. L. Sousa et al. International journal of molecular sciences 2020 sep

Abstract

Cystic fibrosis (CF) cells display a more cancer-like phenotype vs. non-CF cells. KLF4 overexpression has been described in CF and this transcriptional factor acts as a negative regulator of wt-CFTR. KLF4 is described as exerting its effects in a cell-context-dependent fashion, but it is generally considered a major regulator of proliferation, differentiation, and wound healing, all the processes that are also altered in CF. Therefore, it is relevant to characterize the differential role of KLF4 in these processes in CF vs. non-CF cells. To this end, we used wt- and F508del-CFTR CFBE cells and their respective KLF4 knockout (KO) counterparts to evaluate processes like cell proliferation, polarization, and wound healing, as well as to compare the expression of several epithelial differentiation markers. Our data indicate no major impact of KLF4 KO in proliferation and a differential impact of KLF4 KO in transepithelial electrical resistance (TEER) acquisition and wound healing in wt- vs. F508del-CFTR cells. In parallel, we also observed a differential impact on the levels of some differentiation markers and epithelial-mesencymal transition (EMT)-associated transcription factors. In conclusion, KLF4 impacts TEER acquisition, wound healing, and the expression of differentiation markers in a way that is partially dependent on the CFTR-status of the cell.
Aprotinin Inhibits SARS-CoV-2 Replication. D. Bojkova et al. Cells 2020 oct

Abstract

Severe acute respiratory syndrome virus 2 (SARS-CoV-2) is the cause of the current coronavirus disease 19 (COVID-19) pandemic. Protease inhibitors are under consideration as virus entry inhibitors that prevent the cleavage of the coronavirus spike (S) protein by cellular proteases. Herein, we showed that the protease inhibitor aprotinin (but not the protease inhibitor SERPINA1/alpha-1 antitrypsin) inhibited SARS-CoV-2 replication in therapeutically achievable concentrations. An analysis of proteomics and translatome data indicated that SARS-CoV-2 replication is associated with a downregulation of host cell protease inhibitors. Hence, aprotinin may compensate for downregulated host cell proteases during later virus replication cycles. Aprotinin displayed anti-SARS-CoV-2 activity in different cell types (Caco2, Calu-3, and primary bronchial epithelial cell air-liquid interface cultures) and against four virus isolates. In conclusion, therapeutic aprotinin concentrations exert anti-SARS-CoV-2 activity. An approved aprotinin aerosol may have potential for the early local control of SARS-CoV-2 replication and the prevention of COVID-19 progression to a severe, systemic disease.
Genomic determinants implicated in the glucocorticoid-mediated induction of KLF9 in pulmonary epithelial cells. M. M. Mostafa et al. The Journal of biological chemistry 2020 nov

Abstract

Ligand-activated glucocorticoid receptor (GR) elicits variable glucocorticoid-modulated transcriptomes in different cell types. However, some genes, including Kr{\{u}}ppel-like factor 9 (KLF9) a putative transcriptional repressor demonstrate conserved responses. We show that glucocorticoids induce KLF9 expression in the human airways in vivo and in differentiated human bronchial epithelial (HBE) cells grown at air-liquid interface (ALI). In A549 and BEAS-2B pulmonary epithelial cells glucocorticoids induce KLF9 expression with similar kinetics to primary HBE cells in submersion culture. A549 and BEAS-2B ChIP-seq data reveal four common glucocorticoid-induced GR binding sites (GBSs). Two GBSs mapped to the 5'-proximal region relative to KLF9 transcription start site (TSS) and two occurred at distal sites. These were all confirmed in primary HBE cells. Global run-on (GRO)-sequencing indicated robust enhancer RNA (eRNA) production from three of these GBSs in BEAS-2B cells. This was confirmed in A549 cells plus submersion and ALI culture of HBE cells. Cloning each GBS into luciferase reporters revealed glucocorticoid-induced activity requiring a glucocorticoid response element (GRE) within each distal GBS. While the proximal GBSs drove modest reporter induction by glucocorticoids this region exhibited basal eRNA production RNA polymerase II enrichment and looping to the TSS plausibly underlying constitutive KLF9 expression. Post-glucocorticoid treatment interactions between distal and proximal GBSs and the TSS correlated with KLF9 induction. CBP/P300 silencing reduced proximal GBS activity but negligibly effected KLF9 expression. Overall a model for glucocorticoid-mediated regulation of KLF9 involving multiple GBSs is depicted. This work unequivocally demonstrates that mechanistic insights gained from cell-lines can translate to physiologically relevant systems."
Interested in trying STEMCELL's products for respiratory research? Fill out this form to request information about introductory offers.