STEMCELL Technologies IntestiCult IntestiCult Organoid Differentiation Medium (Human)
- 研究用
「IntestiCult™による腸内ウイルス感染の研究」最新情報
IntestiCult™ Organoid Differentiation Medium (Human) (ST-100-0214)は、腸管オルガノイドを3次元(3D)で、あるいは単層または気液界面(ALI)として2次元(2D)で、さらに分化させるための完全培地です。ヒト腸陰窩 (intestinal crypts) に由来する腸管オルガノイド、またはIntestiCult™ Organoid Growth Medium(ヒト用; ST-06010)で継代培養したオルガノイドのいずれかから培養を開始できます。
本品で生成した腸培養物は、分化細胞と幹細胞の集団を生理学的に適切な割合で含み、陰窩-絨毛軸の多様性を再現しています。従来の細胞株と比べて、腸の単層はバリア完全性に優れ、重要な分化マーカーをより高レベルに発現し、in vivoの腸により近い形態を示します。
腸オルガノイド培養の用途には、腸上皮の発達と機能の研究、腸疾患モデル、化合物スクリーニング、および再生医療の開発が含まれます。腸の単層およびALI培養物は頂端面にアクセスしやすいため、特に透過性アッセイと感染症の研究に適しています。
本品の使用前に、腸オルガノイド培養を開始し拡大するには、IntestiCult™ Organoid Growth Medium(ヒト用; ST-06010)が必要になります。
製品の特長
IntestiCult™ ODM (Human) をもちいて、ヒト腸オルガノイドをさらに分化させることができます
- 分化細胞と幹細胞の生理学的比率を維持した腸オルガノイドを生成します
- オルガノイドから単層およびALI培養への移行によって、頂端表面にアクセスしやすくなります
- 継代間および反復間で、一貫した結果が得られます
データ紹介
分化させたヒト腸管オルガノイドの形態
![100-0214-Fig1.jpg](/products/images/100-0214-Fig1_1.jpg)
Figure 1. Differentiated Human Intestinal Organoids Display a Budded Morphology
(A) Organoids grown in IntestiCult™ OGM are primarily cystic.
(B) When switched to IntestiCult™ ODM, organoids develop a thickened epithelium with a pronounced, budded morphology indicative of a more differentiated state. Organoids were imaged on day 5 of expansion or differentiation respectively.
ヒト腸管上皮の成熟細胞マーカーの発現
![100-0214-Fig2.jpg](/products/images/100-0214-Fig2.jpg)
Figure 2. Intestinal Organoids Contain a Higher Proportion of Mature Cell Types Following Differentiation in IntestiCult™ ODM
(A, B) Organoids grown in IntestiCult™ OGM are enriched for Ki-67+ proliferative cells (A), while containing few differentiated cell types such as goblet cells (MUC2, A), enterocytes (KRT20, B), and enteroendocrine cells (CHGA, B).
(C, D) When switched to IntestiCult™ ODM, organoids contain a small number of Ki-67+ proliferative cells (C, arrows), with more physiological proportions of goblet cells (MUC2, C), enterocytes (KRT20, D), and chromogranin A- (CHGA-)positive enteroendocrine cells (D, arrow).
ヒト腸管上皮の気液界面培養における分化
![100-0214-Fig3.jpg](/products/images/100-0214-Fig3_1.jpg)
Figure 3. Differentiation of Intestinal Epithelium at the Air-Liquid Interface (ALI) Using IntestiCult™ ODM
(A – E) Growing organoid-derived monolayers as ALI cultures drives further differentiation of intestinal epithelial cultures as seen by changes in gene expression measured by RT-qPCR. Relative quantification (RQ) for each marker is shown relative to actB and TBP housekeeping genes and normalized with respect to undifferentiated organoids grown in IntestiCult OGM (Human). Progenitor markers (A) Lgr5 and (B) Axin2 are significantly reduced in both submerged monolayers and ALI cultures, while markers of enterocytes (KRT20, C), goblet cells (MUC2, D), and enteroendocrine cells (CHGA, E) are significantly increased. Further reduction in Axin2 is seen in ALI monolayers with an increase in expression of KRT20, MUC2, and CHGA.
(F, G) Comparing cross-sections of organoid monolayers grown in IntestiCult™ ODM as (F) submerged culture or (G) at the ALI shows further differentiation of the intestinal epithelium with an increased proportion of goblet cells and extracellular mucus (MUC2, green).
ヒト腸管オルガノイド由来の単層およびALI培養が示す、より生理的なTEER
![100-0214-Fig4.jpg](/products/images/100-0214-Fig4.jpg)
Figure 4. Differentiated Organoid-Derived Monolayers and ALI Cultures Display More Physiological Trans-Epithelial Electrical Resistance (TEER) than Caco-2 Cells
Differentiated organoid-derived monolayers grown as a submerged monolayer (IntestiCult™ ODM Monolayer), or at the ALI (IntestiCult™ ODM ALI), show higher TEER values as compared to Caco-2 cultures.Organoid-derived monolayers grown at the ALI show a loosening of tight junctions due to further differentiation of the brush border, and thus lower TEER values are observed. * p < 0.0001.
ヒト腸管オルガノイドによるCFTR反応研究モデル
![100-0214-Fig5.jpg](/products/images/100-0214-Fig5.jpg)
Figure 5. Differentiated Intestinal Organoids Provide a Suitable Model for Studying CFTR Response In Vitro
(A) Organoids differentiated further in IntestiCult™ ODM show a comparable degree of swelling when treated with forskolin as compared to organoids grown in IntestiCult™ OGM, demonstrating suitability for use in forskolin-induced swelling assays.
(B – E) Ussing chamber analysis of submerged (B) organoid-derived monolayers and (C) Caco-2 cultures demonstrate increased sensitivity of organoid-derived monolayers to CFTR activation and inhibition by IBMX/Forskolin and CFTR Inhibitor-172 respectively. (D, E) Analysis of CFTR modulation by IBMX/Forskolin and CFTR Inhibitor-172 show significantly greater (D) activation and (E) inhibition of CFTR activity in organoid-derived monolayers as compared to Caco-2 cultures (p < 0.001 for both).