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Updated: Jun 6, 2026
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Robust and Highly Reproducible Generation of Cortical Brain Organoids for Modelling Brain Neuronal Senescence In Vitro
Published on: May 5, 2022
Protocol for minimizing necrotic core formation in iPSC-derived cortical organoids
Md Mahfuz Al Mamun1, Sundari Chetty2
1Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA 02114, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA.
STAR Protocols
|June 4, 2026
Summary
This study presents a novel protocol for creating human cortical organoids (COs) from induced pluripotent stem cells (hiPSCs) without necrotic cores. The method enhances nutrient diffusion, improving long-term viability and structural integrity for neurodevelopment research.
Area of Science:
- Neuroscience
- Developmental Biology
- Stem Cell Biology
Background:
- Human induced pluripotent stem cells (hiPSCs) are used to create cortical organoids (COs) for in vitro modeling of human neurodevelopment.
- Necrotic core formation is a common issue in COs, limiting their long-term viability and utility.
- Current methods to address necrosis often involve organoid slicing, which can damage the structure.
Purpose of the Study:
- To present a protocol for generating hiPSC-derived COs that prevents necrotic core formation.
- To maintain long-term viability and structural integrity of COs without slicing.
- To enhance oxygen and nutrient diffusion within the organoids.
Main Methods:
- Detailed steps for hiPSC culturing and single-cell suspension preparation.
- Procedures for embryoid body (EB) and neuroectoderm formation.
- Methods for CO formation and maintenance focused on preventing necrosis through improved diffusion.
Main Results:
- Successful generation of COs without necrotic core formation.
- Sustained long-term viability of the organoids.
- Preserved structural integrity of the COs due to enhanced diffusion.
Conclusions:
- The developed protocol offers a method to overcome necrotic core formation in hiPSC-derived COs.
- This technique supports extended cultivation and preserves the structural integrity of organoids.
- The protocol is valuable for in vitro modeling of human neurodevelopment.
