製品の特長

PneumaCult™-ALI Mediumをもちいて、ヒト気道上皮細胞を血清・BPE不含条件下で気液界面培養できます

• HBECに広範囲な粘膜繊毛分化を誘導し、ヒト気道に近似した偽重層上皮を形成します
• 血清及びBPE（ウシ下垂体抽出物）フリーのため、変動を最小化できます

ヒト気管支上皮細胞（HBEC）培養の流れ

Phase 1　PneumaCult-Ex Plus（または-Ex）を使用して、液内でヒト気管支上皮細胞（HBEC）を拡大培養します。
Phase 2　ALI培養初期の拡大期では、PneumaCult-Ex Plus（または-Ex）を頂端および基底チャンバーに充填します。
Phase 3　コンフルエンスに達したら両チャンバーから培地を除き、PneumaCult-ALIを基底チャンバーのみに添加することにより、培養物を空気中に引き上げます。21〜28日間のインキュベーション後に偽重層粘液線毛上皮に分化し、1年以上維持できます。

データ紹介

Figure 2. HBECs Cultured in PneumaCult™-Ex Successfully Differentiate into a Pseudostratified Mucociliary Epithelium with PneumaCult™-ALI

Early-passage (P1-3) HBECs cultured in PneumaCult™-Ex successfully differentiate when cultured at air-liquid interface with PneumaCult™-ALI for 28 days. H&E staining revealed the pseudostratifi ed structure of the epithelium with cilia present at the apical surface (A). Periodic acid-Schiff staining demonstrated the presence of goblet cells (B). The presence of ciliated and goblet cells was also demonstrated by immunofl uorescence staining of cilia marker acetylated (AC)-Tubulin (green; C) and the goblet cell marker Mucin5AC (green; D). Appropriate positioning of basal cells along the transwell insert was visualized by immunofl uorescence staining using the basal cell markers p75NTR (green) and p63 (red; E,F). A representative merged image indicates the apical cells, detected by DAPI alone, positioned along the epithelium and in close contact

Figure 3. Electrophysiological characterization of differentiated HBECs (P4) that were expanded in PneumaCult™-Ex Plus, PneumaCult™-Ex, and Bronchial Epithelial Growth Media

TEER (A) and representative characterization of the ion channel activities (B) for ALI cultures at 28 days post air-lift using HBECs expanded in PneumaCult™-Ex Plus, PneumaCult™-Ex, or Bronchial Epithelial Growth Media. Amiloride: ENaC inhibitor. IBMX and Forskolin: CFTR activators. Genistein: CFTR potentiator. CFTRinh-172: CFTR inhibitor. UTP: Calciumactivated Chloride channels (CaCCs) activator. All ALI differentiation cultures were performed using PneumaCult™-ALI.