Mouse Fetal Brain Organoid (Expansion) Kit
MCE Mouse Fetal Brain Organoid (Expansion) Kit contains Mouse Fetal Brain Organoid Expansion Basal Medium and Mouse Fetal Brain Organoid Expansion Culture Supplement . This kit enables the efficient in vitro generation of mouse fetal brain organoids (mFBs). Within this culture system, mouse fetal brain tissue can spontaneously form organoid structures that faithfully recapitulate key features of in vivo cellular heterogeneity and complex tissue organization.
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Storage :
HY-K6143-A: 4°C, 1 year. Shipping with blue ice. HY-K6143-B: -20°C, 1 year. Avoid repeated freeze/thaw cycles. Shipping with dry ice.
- Manual (3641KB)
- COA
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Tel: 609-228-6898 E-mail: [email protected] [email protected]
Description & Advantages
MCE Mouse Fetal Brain Organoid (Expansion) Kit contains Mouse Fetal Brain Organoid Expansion Basal Medium and Mouse Fetal Brain Organoid Expansion Culture Supplement . This kit enables the efficient in vitro generation of mouse fetal brain organoids (mFBs). Within this culture system, mouse fetal brain tissue can spontaneously form organoid structures that faithfully recapitulate key features of in vivo cellular heterogeneity and complex tissue organization.
The culture conditions provided by this kit support long-term and stable expansion of mFBs. The expansion process critically depends on maintaining tissue integrity, which facilitates the formation of a quasi-native extracellular matrix (ECM) microenvironment. This microenvironment effectively promotes sustained proliferation and functional maintenance of the organoids.
This product is suitable for a wide range of applications, including developmental biology research, neurological disease modeling, drug screening, and regenerative medicine. It provides a reliable in vitro platform for studying mouse brain development and elucidating the molecular mechanisms underlying related neurological disorders.
Protocol
Preparation of Complete Culture Medium for Mouse Fetal Brain Organoids
Prepare the complete culture medium according to the table below, mix thoroughly, and keep on ice.
| Reagent | 10 mL | 20 mL | 50 mL | Final concentration |
|---|---|---|---|---|
| Basal Medium A | 9.56 mL | 19.12 mL | 47.8 mL | 1× |
| Supplement B (25×) | 400 μL | 800 μL | 2 mL | 1× |
| Supplement C (250×) | 40 μL | 80 μL | 200 μL | 1× |
Isolation and Generation of Mouse Fetal Brain Organoids
1. Tissue collection: Place freshly isolated mouse fetal brain tissue in ice-cold Tissue Storage Solution (MCE Cat. No.: HY-K6010) and keep temporarily at 4°C. Keep the tissue fully immersed and prevent drying.
2. Tissue pre-treatment: Under sterile conditions and a dissection microscope, remove non-target components such as adipose and muscle tissue with sterile scissors, scalpels, and forceps. Gently rinse with Basal Medium A or PBS until the supernatant is clear and blood/debris are removed.
3. Tissue mincing: Transfer the washed tissue to a sterile dish and cut it into fragments approximately 0.8-1 mm in diameter.
4. Sedimentation and resuspension: Allow fragments to settle for 1-2 min, gently remove the supernatant, resuspend in Basal Medium A or PBS, and repeat the wash once.
5. Distribution: Transfer approximately 20-30 tissue fragments per well into 6-well plates pretreated with Organoid Anti-Adhesion Rinsing Solution (MCE Cat. No.: HY-K6018).
Note: Before transferring fragments, trim approximately 3-5 mm from the pipette tip and keep the pipette vertical to minimize mechanical damage.
6. Organoid culture: Add Mouse Fetal Brain Organoid Complete Medium and culture at 37°C with 5% CO2 on an orbital shaker at 90 rpm.
7. Medium exchange: Replace with freshly prepared, 37°C pre-warmed complete medium every 2-3 d, taking care not to disturb the organoids.
8. Morphology monitoring and passaging: Monitor morphology closely and passage when organoids reach approximately 2-3 mm in diameter.
Organoid Passaging
1. Pre-warm Mouse Fetal Brain Organoid Complete Medium to 37°C.
2. Aspirate the culture supernatant and mechanically fragment the organoids. A recommended method is to resuspend the organoids in 1.5 mL Basal Medium A and cut them into approximately 0.8-1 mm pieces in a sterile dish.
Note: Trim 3-5 mm from the pipette tip before transferring fragments and maintain a vertical pipetting angle to reduce shear stress.
3. Distribute the fragments into HY-K6018-pretreated 6-well plates and gently add pre-warmed complete medium.
4. Continue suspension culture at 37°C and 5% CO2 on an orbital shaker at 90 rpm.
Storage
HY-K6143-A: 4°C, 1 year. Shipping with blue ice.
HY-K6143-B: -20°C, 1 year. Avoid repeated freeze/thaw cycles. Shipping with dry ice.
Attention
1. Tumor cells need to be kept sterile when extracted from primary tumor tissue to avoid contamination from subsequent experiments.
2. This product is for R&D use only, not for drug, household, or other uses.
3. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Cat. No. | Product List | Components | HY-K6143 -100 mL |
HY-K6143 -500 mL |
Storage |
|---|---|---|---|---|---|
| HY-K6143-A | Mouse Fetal Brain Organoid Expansion Basal Medium |
Basal Medium A | 100 mL | 500 mL | 4°C |
| HY-K6143-B | Mouse Fetal Brain Organoid Expansion Culture Supplement |
Culture Supplement B (25×) |
1 mL × 4 | 10 mL × 2 | -20°C. Avoid repeated freeze/thaw cycles. |
| Culture Supplement C (250×) |
0.4 mL | 1 mL × 2 |
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