Organoids can be stained with antibodies for immunocytochemical (ICC)/Immunohistochemical (IHC) analysis using either whole-mount or paraffin embedding/sectioning techniques. It is critical to use antibodies prequalified for organoid analysis.
推薦產品
產品名稱
R-Spondin1 expressing 293T Cell Line, The R-Spondin1-Expressing 293T cell line produces RSPO1, a critical protein used to establish 3D organoid cultures from stomach, small intestine, colon, pancreas, and liver sources.
生物源
human
品質等級
運輸包裝
ambient
一般說明
3D organoid culture systems are increasingly employed as powerful tools for the study of human diseases. Organoids are thought to better represent the tissue-specific niche microenvironment. R-spondin-1 (RSPO1) is one of the most extensively used niche factors for culturing 3D organoids. The first published report of an organoid culture system able to promote long-term survival and multilineage differentiation of intestinal stem cells (ISCs) employed mouse Rspo1 as one of the critical factors [1]. Since then, R-Spondin1 has been used to establish organoid cultures from the stomach, small intestine, colon, pancreas and liver from both mouse and human sources [2] [1].
RSPO1 is an intestinal growth factor that works as a potent activator of the Wnt/beta-catenin signaling pathway by binding to leucine-rich repeat–containing G protein–coupled receptors (LGRs), which are expressed in tissue stem cells. RSPO1 also binds to the transmembrane E3 ubiquitin ligases RNF43 and ZNRF3 and triggers their internalization, thereby potentiating Wnt signaling.
The 293T cell line is stably transfected to express mouse R-spondin1 protein tagged with N-terminus HA epitope tag and C-terminus mouse IgG2α Fc region. R-Spondin1 expressing 293T cell line can be used to produce either purified Rspo1 or Rspo1 conditioned media. The FC region and HA tags enable ease of purification and characterization. Cell line is not the published cell line from the Kuo lab, but rather a related cell line developed in the same lab using the reference protocol.
The 293T cell line is stably transfected to express mouse R-spondin1 protein tagged with N-terminus HA epitope tag and C-terminus mouse IgG2α Fc region. R-Spondin1 expressing 293T cell line can be used to produce either purified Rspo1 or Rspo1 conditioned media. The FC region and HA tags enable ease of purification and characterization. Cell line is not the published cell line from the Kuo lab, but rather a related cell line developed in the same lab using the reference protocol.
細胞系描述
Epithelial Cells
應用
Research Category
Cell Culture
Cell Culture
This product is intended for sale and sold solely for internal non-commercial research use per the terms of the “Restricted Use Agreement” as detailed in the product documentation. For information regarding any other uses, please contact [email protected].
品質
• Each vial contains ≥1X10⁶ viable cells.
• Cells are tested negative for HPV-16, HPV-18, Hepatitis A, B, C, Herpesvirus type 6, 7, 8 and HIV-1 & 2 viruses by PCR.
• Cells are negative for mycoplasma contamination.
• Each lot of cells is genotyped by STR analysis to verify the unique identity of the cell line.
• Cells are tested negative for HPV-16, HPV-18, Hepatitis A, B, C, Herpesvirus type 6, 7, 8 and HIV-1 & 2 viruses by PCR.
• Cells are negative for mycoplasma contamination.
• Each lot of cells is genotyped by STR analysis to verify the unique identity of the cell line.
儲存和穩定性
Store in liquid nitrogen. The cells can be cultured for at least 10 passages after initial thawing without significantly affecting the cell marker expression and functionality.
免責聲明
RESEARCH USE ONLY. This product is regulated in France when intended to be used for scientific purposes, including for import and export activities (Article L 1211-1 paragraph 2 of the Public Health Code). The purchaser (i.e. enduser) is required to obtain an import authorization from the France Ministry of Research referred in the Article L1245-5-1 II. of Public Health Code. By ordering this product, you are confirming that you have obtained the proper import authorization.
儲存類別代碼
10 - Combustible liquids
水污染物質分類（WGK）
WGK 1
閃點（°F）
Not applicable
閃點（°C）
Not applicable
分析證明 (COA)
輸入產品批次/批號來搜索 分析證明 (COA)。在產品’s標籤上找到批次和批號，寫有 ‘Lot’或‘Batch’.。
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文章
器官組織培養常見問題。瞭解如何按照既定的 3D 細胞培養方案培養和分析器官組織。技巧與竅門。
Organoid culture FAQs. Learn how to culture and analyze organoids following established 3D cell culture protocols. Tips and tricks.
類器官培養產品，用於產生組織和幹細胞衍生的 3D 大腦、腸道、腸道、肺和癌症腫瘤類器官模型。
Organoid culture products to generate tissue and stem cell derived 3D brain, intestinal, gut, lung and cancer tumor organoid models.
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How do you stain organoids with antibodies?
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How Do you Extract RNA/DNA from Organoids in Cultured in Matrigel?
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Traditional DNA/RNA isolation kits can be used to prepare genomic material for sequencing (RNA-seq, scRNA-seq, WGS, CHIP-seq etc). For some applications with large organoids, tissue specific DNA/RNA extraction kits are recommended. TRI Reagent (T9424) can be added to the dissociated organoids before RNA extraction and can be stored at -80 °C until future use.
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How are organoids cultured?
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Once isolated, organoids are cultured in specialized media embedded in a rich extracellular matrix (ECM) basement membrane extract hydrogel such as Growth Factor Reduced Matrigel. Media is replenished every other day and cells are passaged once per week (7-12 days) before cells become too large or necrotic. Cells can be passaged in either small clump fragments or as single cells using mechanical dissociation/enzyme-free passaging reagents. Depending on organoid type and confluency, split ratios of roughly 1:3-1:4 can be used when passaging organoids. For sensitive organoid types, ROCKi (Y27632) (SCM075) can be added during passaging to help in promoting cell viability.
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What is the selection marker that is used in your RS Spondin expressing 293 cells. Is it neomycin resistance? What antibiotic should be used for selection pressure? Geneticin/G418?
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The cell line is selected by zeocin. It is not necessary to add zeocin to culture medium as the cell line is stable.
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