产品号 #100-0384_C
用于从原代肝组织生成类器官的培养液
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Reagent for counting viable mammalian cells
Reagent for lysis of red blood cells
Clear polystyrene flat-bottom, tissue culture-treated multiwell cell culture plate with lid
RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2
Compatible antibodies for purity assessment of isolated cells
使用 HepatiCult™ 类器官起始培养基(OIM)(人源)可高效生成具有生理相关性的人源肝脏类器官,适用于不同供体来源。
HepatiCult™ OIM 既可单独购买,也可作为 HepatiCult™ 类器官试剂盒(人源)的一部分,用于从新鲜或冷冻保存的人肝组织中生成肝脏类器官。这些类器官可在 HepatiCult™ 类器官生长培养基(人源)中进一步扩增和维持,随后使用 HepatiCult™ 类器官分化培养基(人源)进行分化,获得具有肝脏功能(包括 CYP3A4 活性)的成熟类器官。
如需了解更多关于 HepatiCult™ 类器官培养方案的信息,请参阅技术手册和教育资料。
如果您打算将此产品用于商业目的,请通过www.huborganoids.nl联系HUB以获得商业使用许可或有关HUB许可的说明。
HepatiCult™ OIM 既可单独购买,也可作为 HepatiCult™ 类器官试剂盒(人源)的一部分,用于从新鲜或冷冻保存的人肝组织中生成肝脏类器官。这些类器官可在 HepatiCult™ 类器官生长培养基(人源)中进一步扩增和维持,随后使用 HepatiCult™ 类器官分化培养基(人源)进行分化,获得具有肝脏功能(包括 CYP3A4 活性)的成熟类器官。
如需了解更多关于 HepatiCult™ 类器官培养方案的信息,请参阅技术手册和教育资料。
如果您打算将此产品用于商业目的,请通过www.huborganoids.nl联系HUB以获得商业使用许可或有关HUB许可的说明。
HepatiCult™ OIM 既可单独购买,也可作为 HepatiCult™ 类器官试剂盒(人源)的一部分,用于从新鲜或冷冻保存的人肝组织中生成肝脏类器官。这些类器官可在 HepatiCult™ 类器官生长培养基(人源)中进一步扩增和维持,随后使用 HepatiCult™ 类器官分化培养基(人源)进行分化,获得具有肝脏功能(包括 CYP3A4 活性)的成熟类器官。
如需了解更多关于 HepatiCult™ 类器官培养方案的信息,请参阅技术手册和教育资料。
如果您打算将此产品用于商业目的,请通过www.huborganoids.nl联系HUB以获得商业使用许可或有关HUB许可的说明。
HepatiCult™ OIM 既可单独购买,也可作为 HepatiCult™ 类器官试剂盒(人源)的一部分,用于从新鲜或冷冻保存的人肝组织中生成肝脏类器官。这些类器官可在 HepatiCult™ 类器官生长培养基(人源)中进一步扩增和维持,随后使用 HepatiCult™ 类器官分化培养基(人源)进行分化,获得具有肝脏功能(包括 CYP3A4 活性)的成熟类器官。
如需了解更多关于 HepatiCult™ 类器官培养方案的信息,请参阅技术手册和教育资料。
如果您打算将此产品用于商业目的,请通过www.huborganoids.nl联系HUB以获得商业使用许可或有关HUB许可的说明。
亚型
专用培养基
细胞类型
肝细胞
种属
人
应用
类器官培养
品牌
HepatiCult
研究领域
疾病建模,药物发现和毒理检测,上皮细胞研究,类器官,干细胞生物学
制剂类别
无血清
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.
Table 1. Product Recommendations for Liver Organoid Initiation, Expansion, and Differentiation
The recommended configuration of the HepatiCult™ Organoid Kit (Human) may differ based on starting material and experimental goals. When establishing liver organoid cultures from human liver tissue, HepatiCult™ Organoid Initiation Medium (OIM; Human) is recommended for efficient organoid initiation (see Figure 2 below). The expansion of already established organoids (fresh in culture or cryopreserved) is supported by HepatiCult™ Organoid Growth Medium (OGM; Human). These organoids should be maintained for 2-3 passages before further differentiation using HepatiCult™ Organoid Differentiation Medium (ODM). Refer to the Product Manual (Document #10000008301) for full culturing protocols.
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).
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.
Figure 4. Expansion of Organoid Cultures in HepatiCult™ Organoid Growth Medium
Hepatic organoids show efficient growth in HepatiCult™ OGM across multiple donors and passages with potential for indefinite culture. Organoids initiated using the serum-free workflow exhibited comparable growth rates when expanded in HepatiCult™ OGM (open markers).
Figure 5. 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.
Figure 6. 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).
Figure 8. Differentiation of Hepatic Organoids in HepatiCult™ ODM Induces Changes in Gene Expression Consistent With Hepatic Maturation
Upon differentiation in HepatiCult™ ODM, liver organoids show changes in gene expression with a decrease in expression of stem cell marker LGR5, as well as of ductal markers CK19, CK7, and SOX9, and an increase in expression of hepatic markers HNF4A, ALB, CYP3A4, and ASGR1. Each replicate represents an individual donor sample analyzed by qPCR at passage 4 on day 8 in HepatiCult™ OGM and day 15 in HepatiCult™ ODM, showing relative gene expression levels normalized to TBP and UBC housekeeping genes.
Figure 9. Differentiated Hepatic Organoids Demonstrate Functionality of Mature Hepatocytes
Upon differentiation in HepatiCult™ ODM, liver organoids were assayed for (A) albumin secretion, (B) CYP3A4 activity, (C) total bile acid production, and (D) urea production. Hepatic functionalities were compared to HepG2 cells and primary human hepatocytes (PHH), which were cultured in supplier-recommended media. Albumin secretion was detected using an ELISA kit (Abcam), total bile acid and urea production were analysed using colorimetric kits (Abcam), and CYP43A4 activity, referring to baseline activity without induction, was determined using the Luciferin-IPA kit (Promega). (mean ± SD; n = 3 organoid lines across 2 experiments, n = 2-3 technical replicates of HepG2 in 1 experiment, and n = 3 cryopreserved PHH donor samples in 1 experiment), * p < 0.05; ** p < 0.01.
Figure 10. HepatiCult™ Organoid Kit Supports the Growth and Differentiation of Porcine Liver Organoids
Porcine liver organoids were established in HepatiCult™ Organoid Initiation Medium, and subsequently expanded for 4 passages in (A) HepatiCult™ Organoid Growth Medium (Human) and differentiated in (B, C) HepatiCult™ Organoid Differentiation Medium (Human). Organoids are stained with basolateral protein marker p120 (green), apical membrane and bile canaliculi marker F-Actin (purple), and nuclear dye Hoechst (blue). (D) Magnification of Panel C indicates bile canaliculi formation as seen through F-Actin staining. Scale bars = 50 μm. Data used with permission from Dr. Amy Engevik (Vanderbilt University Medical Center)
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种属 | Human |
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配方类别 | Serum-Free |
用于建立和维持小鼠肝祖类器官的培养基
用于将人多能干细胞分化为肝细胞样细胞的无血清分化试剂盒
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