제품특징
HepatiCultTM Organoid kit (Human)은 인간 간 오가노이드를 안정적으로 생성, 확장 및 분화를 도와주는 키트 입니다. 키트는 신선하거나 냉동 보존된 인간 간 조직에서 오가노이드를 배양하는데 필요한 모든 구성 요소가 포함되어 있습니다.
Advantages
- 인간 간 오가노이드의 확립, 유지, 분화를 위한 완전한 배양 시스템 제공
- 다양한 간 조직에서 효율적인 오가노이드 개시 가능
- CYP3A4 활성을 나타내는 성숙한 간 오가노이드 생성 가능
General Workflow
Figure 1. HepatiCult™ Organoid Kit (Human) Enables Liver Organoid Initiation, Expansion, and Differentiation
Human liver organoids can be grown from normal human liver tissue-derived hepatic ducts using the HepatiCult™ Organoid Kit (Human). (A) Cultures are established in HepatiCult™ Organoid Initiation Medium (OIM; Human) or HepatiCult™ Organoid Growth Medium (OGM; Human) (see Table 1 below) and subsequently passaged in HepatiCult™ OGM for expansion. (B) After passaging 2-3 times in HepatiCult™ OGM, cultures can be switched to HepatiCult™ Organoid Differentiation Medium (ODM; Human) to differentiate organoids towards more mature hepatic cell types. Refer to the Product Manual (Document #10000008301) for full culturing protocols.
효율적인 오가노이드 생성 가능
Figure 2. HepatiCult™ Organoid Kit (Human) Provides Efficient Organoid Initiation From Human Liver Tissue
(A) Organoid cultures were initiated in HepatiCult™ OIM, and then (B) passaged into HepatiCult™ OGM. For serum-free culture conditions, organoid cultures were both, (C) initiated in and (D) passaged in HepatiCult™ OGM. Culture images shown are from (A, C) day 15 following initiation, and (B, D) on day 8 of the first passage. (E) Quantification of organoid initiation efficiency shows a significantly higher organoid yield in HepatiCult™ OIM per gram of human liver tissue (mean ± SD; n=14).
다양한 간 조직에서 오가노이드 생성 가능
Figure 3. HepatiCult™ Organoid Initiation Medium (Human) Supports Robust Organoid Establishment Across Multiple Liver Tissue Donor Samples
Organoids initiated from 4 donor tissue samples (A-D) exhibit morphological heterogeneity 15 days after initiation. All initiated cultures were subsequently expanded in HepatiCult™ OGM using a 1:1 passaging ratio (E-H), yielding healthy organoids at the end of the first passage.
Hepatic 오가노이드의 progenitor 확인 가능
Figure 4. Proliferating Hepatic Organoids Display Characteristics of Hepatic Progenitors
Human liver organoids grown in HepatiCult™ OGM display characteristics of proliferating hepatic progenitors observed through immunocytochemistry staining of (A) KI67, (B) HNF4A and (C) SOX9. Proliferating hepatic organoids also display characteristics of the hepatic epithelium including expression of (A) EPCAM. (B, C) Nuclei are counterstained with DAPI.
오가노이드 계대 간의 stem cell 마커 유지
Figure 5. Proliferating Hepatic Organoids Maintain Genetic Expression Across Multiple Passages
Liver organoids maintained in HepatiCult™ OGM express stem cell markers (A) LGR5 and (B) AXIN2, ductal markers (C) SOX9 and (D) KRT19, as well as hepatic marker (E) HNF4a and (F) Albumin (ALB) across multiple passages, with minimal albumin expression observed during culture in HepatiCult™ OGM. Expression levels were measured by qPCR and normalized to TBP and UBC housekeeping genes to quantify relative expression levels. (mean ± SD; n = 2-5 organoid lines), * p < 0.05.
Figure 7. Organoid Differentiation Induces Changes in Organoid Morphology
Organoids exhibit a compact and dense morphology, often comprising thickened epithelia, upon switching cultures to HepatiCult™ Organoid Differentiation Medium (ODM). Shown are images of the same culture well over the course of the differentiation, including (A) day two of culture in HepatiCult™ OGM, (B) day five of culture, immediately after switching organoid cultures from HepatiCult™ OGM to HepatiCult™ ODM, (C) day seven of culture (two days after switching to ODM), (D) day ten of culture (five days after switching to ODM), and (E) day 15 of culture (ten days after switching to ODM). (F) Magnification of the rectangular section highlighted in (E).
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