PneumaCult™-ALI-S介质

在气液界面培养的人小气道上皮细胞的无血清和无bpe培养基

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产品号 #05050_C

在气液界面培养的人小气道上皮细胞的无血清和无bpe培养基

产品优势

在PneumaCult™-ALI-S中培养的人小气道上皮细胞（HSAEC）经过广泛的粘膜纤毛分化，形成与小气道上皮非常相似的立方上皮
PneumaCult™-ALI-S不含血清和bpe，可最大限度地减少差异
PneumaCult™-ALI-S和PneumaCult™-Ex Plus构成了一个完整的、优化的系统，用于扩展、维护和区分HSAEC

产品组分包括

PneumaCult™-ALI-S基础培养基，450毫升
PneumaCult™-ALI-S补充剂（10X）， 50 mL
PneumaCult™-ALI-S维护补充剂（100X）， 5 x 1ml

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概述

PneumaCult™-ALI- s Medium（目录#05050）是一种不含血清和bpe的培养基，用于在气液界面（ALI）培养人小气道上皮细胞。在PneumaCult™-ALI-S培养基中培养的小气道上皮细胞经过广泛的粘膜纤毛分化形成立方体上皮，其形态和功能特征与体内的人小气道相似。

一起，PneumaCult™- ari - s Medium和PneumaCult™-Ex Plus Medium（#05040）构成了一个完全集成的无bpe培养系统，用于体外人体小气道建模。这一强大而明确的系统是基础呼吸研究、毒性研究和药物开发的宝贵工具。

了解如何培养人类气道上皮细胞在我们的ALI按需肺课程或浏览我们的常见问题（FAQs）关于使用PneumaCult™的ALI培养工作流程。

Data Figures

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.

Protocols and Documentation

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

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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

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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

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.

Publications (1)

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.

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