Human iPSC/ESC Cortical Brain Organoid Maturation Medium 

MCE Human iPSC/ESC Cortical Brain Organoid Maturation Medium is a ready-to-use culture system specifically formulated for the long-term maintenance and maturation of cortical brain organoids. It serves as a seamless transition from the induction phase and supports the continued growth and functional development of the organoids. Upon switching PSC-derived cortical brain organoids to this medium, organoid viability and structural stability are markedly enhanced, enabling a robust and extended culture period of up to 180 d.

1. DPBS (MCE Cat. No.: HY-K3008)

2. Basement Membrane Matrix IPSC-qualified (MCE Cat. No.: HY-K6006)

3. iPSC/ESC Dissociation Solution (Enzyme-Free) (MCE Cat. No.: HY-K6020)

4. CEPT Cocktail Plus (1000×) (MCE Cat. No.: HY-K6021)

5. ECM Gentle Dissociation Solution (MCE Cat. No.: HY-K6022)

6. Human iPSC/ESC Cortical Brain Organoid Induction Differentiation Kit (MCE Cat. No.: HY-K6301)

7. Human PSC Maintenance Medium (MCE Cat. No.: HY-K6501)

8. Equipment and Consumables: 6/96-well low-attachment plates, and multichannel pipettes (8-12 channels), among other related equipment and consumables.

Human iPSC/ESC Cortical Brain Organoid Induction Differentiation Kit (MCE Cat. No.: HY-K6301) can theoretically generate no fewer than 100 brain organoids. For the initial induction, it is recommended to start with one 6-well plate, which typically yields approximately 60-100 uniformly sized neuroepithelial spheres (ideal formation rate ~60%).

1. (Refer to the HY-K6301 Manual) Neuroepithelial Sphere Induction and Neural Rosette Expansion

Using the Human iPSC/ESC Cortical Brain Organoid Induction Differentiation Kit (MCE Cat. No.: HY-K6301), PSCs are induced to form neuroepithelial spheres and initiate neural rosette expansion. When black punctate structures appear in the center and periphery of the spheres, indicating well-developed neural rosettes, discard the expansion medium completely.

2. Transition to the Maturation Stage

Add 2 mL of Human iPSC/ESC Cortical Brain Organoid Maturation Medium to each well and transfer the plate to a cross-shaker/rotary shaker at 80 rpm for 18-26 d, refreshing the medium every 2-3 d. Mature brain organoids at this stage are suitable for characterization and downstream applications. When cultured long-term on a shaker using the maturation medium, organoids can be maintained up to 120 d without central apoptosis.

Note: Mature organoids exhibit layered VZ- and SVZ-like structures composed of TUJ1⁺ neurons and SOX2⁺ radial glia, as confirmed by immunofluorescence (IF). qPCR analysis shows a 3-5-fold increase in glutamatergic markers such as SLC17A6/VGLUT2.

3. (Refer to the HY-K6301 Manual) Organoid Sectioning

When cortical organoids exceed 1 mm in diameter, perform mechanical sectioning to alleviate inner-core apoptosis. After cutting, change the medium daily to remove apoptosis-related debris and continue shaking culture at 80 rpm for 4 d.

4. Long-Term Maintenance

After 4 d, switch to the Cortical Brain Organoid Maturation Medium and maintain the organoids at 37°C, 120 rpm. This regimen supports extended culture and organoid viability for up to 180 d.

Note: During culture, organoid quality can be monitored by TUNEL assay for central necrosis and by evaluating markers such as Synaptophysin to assess maturation and functional status.

1. This product is sterile and should be handled using aseptic techniques. It is recommended to aliquot and store to avoid contamination.

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.