Mouse Intestinal Organoid Plus Kit
MCE Mouse Intestinal Organoid Plus Kit is an upgraded serum-free culture system developed based on the Mouse Small Intestinal Organoid Culture Kit (HY-K6119), designed for the establishment, expansion, and long-term maintenance of mouse small intestinal organoids. The kit contains basal culture medium and multiple essential supplements, and an optional Wnt ligand supplement is provided to enable modulation of Wnt signaling activity in the culture system.
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Storage :
Basal Medium A: 4°C, 1 year. Shipping with blue ice.
Supplement B (25×), Supplement C (100×), Supplement D (Wnt/β-catanin ligand): -20°C, 1 year. Avoid repeated freeze/thaw cycles. Shipping with dry ice.
Supplement E (EDTA): RT, 1 year. Shipping with blue ice.
- Manual (5995KB)
- COA
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
Description & Advantages
MCE Mouse Intestinal Organoid Plus Kit is an upgraded serum-free culture system developed based on the Mouse Small Intestinal Organoid Culture Kit (HY-K6119), designed for the establishment, expansion, and long-term maintenance of mouse small intestinal organoids. The kit contains basal culture medium and multiple essential supplements, and an optional Wnt ligand supplement is provided to enable modulation of Wnt signaling activity in the culture system.
This product is based on the self-renewal and multilineage differentiation capacity of stem cells and progenitor cells located at the base of intestinal crypts, allowing the stable generation and maintenance of mouse small intestinal organoids with typical architecture and cellular composition. The organoids recapitulate key physiological features of native small intestinal tissue and are suitable for studies of intestinal development, physiological functions, disease mechanisms, and drug screening.
The Wnt supplement can be selectively added during the initial organoid establishment phase or during single-cell passaging to enhance crypt budding, maintain stemness, and improve organoid formation efficiency and viability. In models with Paneth cell deficiency or limited endogenous Wnt signaling, supplementation with the Wnt ligand effectively supports early organoid formation.
Once stable organoid structures are established, withdrawal of the Wnt supplement promotes differentiation toward mature small intestinal phenotypes, resulting in organoids composed of multiple intestinal cell types, including LGR5+ intestinal stem cells, KI67+ proliferating cells, ALPi+ absorptive enterocytes, ATOH1+ Paneth cells, and MUC2+ goblet cells.
Protocol
1. Preparation of Mouse Intestinal Organoid Plus Complete Medium
| Reagent | 10 mL complete medium |
|---|---|
| Mouse Intestinal Organoid Plus Basal Medium A | 9.5 mL (without optional Supplement D) |
| Organoid Culture Supplement B (25×) | 400 μL |
| Organoid Culture Supplement C (100×) | 100 μL |
| Organoid Culture Supplement D | Optional; add according to experimental design or Manual |
Mix thoroughly and keep on ice. Organoid Culture Supplement D is an optional Wnt/β-catenin ligand-related component and may be added according to the experimental purpose.
2. Isolation of Small-Intestinal Crypts
a. Collect fresh small intestine, rinse with pre-cooled PBS to remove luminal contents, open longitudinally, cut into small pieces, and wash repeatedly.
b. Prepare a crypt-release working solution using Supplement E (EDTA) supplied with the kit and incubate the tissue until crypts begin to loosen.
c. Remove the EDTA working solution, add pre-cooled PBS, release crypts by shaking or gentle pipetting, and remove large tissue debris using an appropriate cell strainer.
d. Centrifuge at 250-300 × g for 3-5 min, collect the crypt pellet, and wash 1-2 times with Basal Medium A.
3. Construction of Organoid
a. Resuspend crypts in MCE Basement Membrane Matrix on ice.
b. Dispense approximately 25-35 μL per well into a 24-well plate, avoiding bubbles. Incubate at 37°C with 5% CO2 for 15-30 min until gelled.
c. Slowly add 500 μL Mouse Intestinal Organoid Plus complete medium per well.
d. Replace with fresh medium every 2-3 days. Depending on the experimental purpose, compare organoid growth with or without Supplement D.
4. Organoid Passaging
a. When organoids enlarge or become overly dense, aspirate the medium and detach the matrix/organoids from the well.
b. Mechanically pipette or briefly dissociate the organoids into smaller clusters while avoiding over-digestion.
c. Centrifuge at 250-300 × g for 3-5 min, discard the supernatant, and wash.
d. Re-embed on ice in Basement Membrane Matrix at 25-35 μL per well, allow the matrix to gel, add fresh complete medium, and continue culture.
Storage
Basal Medium A: 4°C, 1 year. Shipping with blue ice.
Supplement B (25×), Supplement C (100×), Supplement D (Wnt/β-catanin ligand): -20°C, 1 year. Avoid repeated freeze/thaw cycles. Shipping with dry ice.
Supplement E (EDTA): RT, 1 year. Shipping with blue ice.
Attention
1. Primary cells need to be kept sterile when extracted from primary tissue to avoid contamination from subsequent experiments.
2. Operations involving MCE Basement Membrane Matrix need to be kept at low temperature throughout. MCE Basement Membrane Matrix should be injected rapidly into the cell culture wells after resuspension with the cells, while avoiding air bubbles.
3. This product is for R&D use only, not for drug, household, or other uses.
4. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Cat. No. | Product List | Components | HY-K6119P-100 mL | HY-K6119P-500 mL | Storage |
|---|---|---|---|---|---|
| HY-K6119P-A | Mouse Intestinal Organoid Basal Medium |
Basal Medium A | 100 mL | 500 mL | 4°C, 1 year. |
| HY-K6119P-B | Mouse Intestinal Organoid Culture Supplement 1 |
Supplement B (25×) | 1 mL × 2 | 10 mL | -20°C, 1 year. Avoid repeated freeze/thaw cycles. |
| Supplement C (100×) | 0.4 mL | 1 mL × 2 | |||
| Supplement D (Wnt/β-catanin ligand) |
0.4 mL | 1 mL × 2 | |||
| HY-K6119P-C | Mouse Intestinal Organoid Culture Supplement 2 |
Supplement E (EDTA) | 0.2 mL | 1 mL | RT, 1 year. |
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