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Protocol for measuring lipid membrane fluidity in human iPSC-derived neural cells.
Satoshi Morita1, Takayuki Kondo2, Haruhisa Inoue2
1iPSC-based Drug Discovery and Development Team, RIKEN BioResource Research Center (BRC), Kyoto 619-0237, Japan; Institute for Science of Life, Suntory Wellness Ltd., Kyoto 619-0284, Japan; Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
This study details a protocol for differentiating human cortical neurons from induced pluripotent stem cells (iPSCs). The method enables analysis of lipid membrane fluidity, crucial for understanding neuronal function and disease.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Biochemistry
Background:
- Induced pluripotent stem cells (iPSCs) offer a valuable model for studying human neuronal development and function.
- Lipid membrane fluidity is a critical biophysical property influencing neuronal signaling and health.
- Understanding neuronal differentiation and membrane properties is essential for neurodegenerative disease research.
Purpose of the Study:
- To present a detailed protocol for differentiating human cortical neurons from iPSCs.
- To describe a method for analyzing lipid membrane fluidity in these differentiated neurons.
- To facilitate the investigation of how membrane fluidity impacts neuronal function and pathophysiology.
Main Methods:
- Protocol for embryoid body formation from iPSCs.
- Neuronal induction and adherent culture maturation techniques.
- Live-cell measurement of lipid membrane fluidity using the LipiORDER fluorescent probe.
Main Results:
- Successful differentiation of human cortical neurons from iPSCs.
- Quantification of lipid membrane fluidity in mature neurons.
- Establishment of a method linking neuronal differentiation to membrane biophysics.
Conclusions:
- The presented protocol provides a robust method for generating human cortical neurons from iPSCs.
- This approach allows for the examination of lipid membrane fluidity's role in neuronal physiology and disease.
- The study enables future investigations into therapeutic strategies targeting membrane properties.
