PneumaCult™-Ex Medium

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

More Views

PneumaCult™-Ex Medium

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

500 mL
Catalog #05008
146 USD

Required Products

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.
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
Components:
  • PneumaCult™-Ex Basal Medium, 490 mL
  • PneumaCult™-Ex 50X Supplement, 10 mL
Subtype:
Specialized Media
Cell Type:
Airway Cells
Species:
Human
Application:
Cell Culture; Expansion; Maintenance
Brand:
PneumaCult
Area of Interest:
Epithelial Cell Biology
Formulation:
Serum-Free

Scientific Resources

Educational Materials

(9)

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

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).

Publications

(9)
International journal of cancer 2018 JUL

Conditionally reprogrammed cells (CRC) methodology does not allow the in vitro expansion of patient-derived primary and metastatic lung cancer cells.

G. Sette et al.

Abstract

Availability of tumor and non-tumor patient-derived models would promote the development of more effective therapeutics for non-small cell lung cancer (NSCLC). Recently, conditionally reprogrammed cells (CRC) methodology demonstrated exceptional potential for the expansion of epithelial cells from patient tissues. However, the possibility to expand patient-derived lung cancer cells using CRC protocols is controversial. Here, we used CRC approach to expand cells from non-tumoral and tumor biopsies of patients with primary or metastatic NSCLC as well as pulmonary metastases of colorectal or breast cancers. CRC cultures were obtained from both tumor and non-malignant tissues with extraordinary high efficiency. Tumor cells were tracked in vitro through tumorigenicity assay, monitoring of tumor-specific genetic alterations and marker expression. Cultures were composed of EpCAM+ lung epithelial cells lacking tumorigenic potential. NSCLC biopsies-derived cultures rapidly lost patient-specific genetic mutations or tumor antigens. Similarly, pulmonary metastases of colon or breast cancer generated CRC cultures of lung epithelial cells. All CRC cultures examined displayed epithelial lung stem cell phenotype and function. In contrast, brain metastatic lung cancer biopsies failed to generate CRC cultures. In conclusion, patient-derived primary and metastatic lung cancer cells were negatively selected under CRC conditions, limiting the expansion to non-malignant lung epithelial stem cells from either tumor or non-tumor tissue sources. Thus, CRC approach cannot be applied for direct therapeutic testing of patient lung tumor cells, as the tumor-derived CRC cultures are composed of (non-tumoral) airway basal cells.
Journal of cystic fibrosis : official journal of the European Cystic Fibrosis Society 2018 APR

The CF Canada-Sick Kids Program in individual CF therapy: A resource for the advancement of personalized medicine in CF.

P. D. W. Eckford et al.

Abstract

BACKGROUND Therapies targeting certain CFTR mutants have been approved, yet variations in clinical response highlight the need for in-vitro and genetic tools that predict patient-specific clinical outcomes. Toward this goal, the CF Canada-Sick Kids Program in Individual CF Therapy (CFIT) is generating a first of its kind"
Biology open 2018 APR

Fank1 and Jazf1 promote multiciliated cell differentiation in the mouse airway epithelium.

J.-A. Johnson et al.

Abstract

The airways are lined by secretory and multiciliated cells which function together to remove particles and debris from the respiratory tract. The transcriptome of multiciliated cells has been extensively studied, but the function of many of the genes identified is unknown. We have established an assay to test the ability of over-expressed transcripts to promote multiciliated cell differentiation in mouse embryonic tracheal explants. Overexpression data indicated that Fibronectin type 3 and ankyrin repeat domains 1 (Fank1) and JAZF zinc finger 1 (Jazf1) promoted multiciliated cell differentiation alone, and cooperatively with the canonical multiciliated cell transcription factor Foxj1. Moreover, knock-down of Fank1 or Jazf1 in adult mouse airway epithelial cultures demonstrated that these factors are both required for ciliated cell differentiation in vitro This analysis identifies Fank1 and Jazf1 as novel regulators of multiciliated cell differentiation. Moreover, we show that they are likely to function downstream of IL6 signalling and upstream of Foxj1 activity in the process of ciliated cell differentiation. In addition, our in vitro explant assay provides a convenient method for preliminary investigation of over-expression phenotypes in the developing mouse airways.This article has an associated First Person interview with the first author of the paper.
Pulmonary pharmacology & therapeutics 2017 JUN

Neutralization of both IL-1α/IL-1β plays a major role in suppressing combined cigarette smoke/virus-induced pulmonary inflammation in mice.

Bucher H et al.

Abstract

Smoking is an important risk factor for the development of chronic obstructive pulmonary disease (COPD) and viral infections are believed to be major triggers of exacerbations, which periodically lead to a worsening of symptoms. The pro-inflammatory IL-1 family members IL-1α and IL-1β are increased in COPD patients and might contribute to disease pathology. We investigated whether individual or combined inhibition of these cytokines reduced lung inflammation in cigarette smoke (CS)-exposed and H1N1-infected BALB/c mice. Animals were treated with individual or combined antibodies (Abs) directed against IL-1α, IL-1β or IL-1R1. Cells in BAL fluid and cytokines/chemokines in lung homogenate were determined. The viral load was investigated. Blocking IL-1α had significant suppressive effects on total cells, neutrophils, and macrophages. Furthermore, it reduced KC levels significantly. Blocking of IL-1β did not provide significant activity. In primary human bronchial epithelial air-liquid-interface cell cultures infected with H1N1, IL-1α Abs but not IL-1β Abs reduced levels of TNF-α and IL-6. Concomitant usage of Abs against IL-1α/IL-1β revealed strong effects in vivo and reduced total cells, neutrophils and macrophages. Additionally, levels of KC, IL-6, TNF-α, MCP-1, MIP-1α and MIP-1β were significantly reduced and ICAM-1 and MUC5 A/C mRNA expression was attenuated. The viral load decreased significantly upon combined IL-1α/IL-1β Ab treatment. Blocking the IL-1R1 provided significant effects on total cells, neutrophils and macrophages but was inferior compared to inhibiting both its soluble ligands IL-1α/IL-1β. Our results suggest that combined inhibition of IL-1α/IL-1β might be beneficial to reduce CS/H1N1-induced airway inflammation. Moreover, combined targeting of both IL-1α/IL-1β might be more efficient compared to individual neutralization IL-1α or IL-1β or inhibition of the IL-1R1.
Scientific reports 2017 FEB

Novel flow cytometry approach to identify bronchial epithelial cells from healthy human airways.

Maestre-Batlle D et al.

Abstract

Sampling various compartments within the lower airways to examine human bronchial epithelial cells (HBEC) is essential for understanding numerous lung diseases. Conventional methods to identify HBEC in bronchoalveolar lavage (BAL) and wash (BW) have throughput limitations in terms of efficiency and ensuring adequate cell numbers for quantification. Flow cytometry can provide high-throughput quantification of cell number and function in BAL and BW samples, while requiring low cell numbers. To date, a flow cytometric method to identify HBEC recovered from lower human airway samples is unavailable. In this study we present a flow cytometric method identifying HBEC as CD45 negative, EpCAM/pan-cytokeratin (pan-CK) double-positive population after excluding debris, doublets and dead cells from the analysis. For validation, the HBEC panel was applied to primary HBEC resulting in 98.6% of live cells. In healthy volunteers, HBEC recovered from BAL (2.3% of live cells), BW (32.5%) and bronchial brushing samples (88.9%) correlated significantly (p = 0.0001) with the manual microscopy counts with an overall Pearson correlation of 0.96 across the three sample types. We therefore have developed, validated, and applied a flow cytometric method that will be useful to interrogate the role of the respiratory epithelium in multiple lung diseases.
STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.