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Isolation and Expansion of Neurospheres from Postnatal (P1−3) Mouse Neurogenic Niches
Published on: May 23, 2020
A method for efficiently generating neurospheres from human-induced pluripotent stem cells using microsphere arrays.
Tomoko Shofuda1, Hayato Fukusumi, Daisuke Kanematsu
1Division of Stem Cell Research, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
Neuroreport
|December 15, 2012
Summary
Researchers developed a new method using microsphere arrays to efficiently generate and propagate human induced pluripotent stem cell-derived neural progenitor cells (hiPSC-NSPCs) in neurospheres. This advance facilitates their use in regenerative medicine and neurotoxicity studies.
Area of Science:
- Stem Cell Biology
- Neuroscience
- Regenerative Medicine
Background:
- Human neural stem and progenitor cells (hN-NSPCs) are crucial for research but difficult to obtain.
- Human embryonic stem cell-derived (hESC-NSPCs) and human induced pluripotent stem cell-derived neural progenitor cells (hiPSC-NSPCs) are viable alternatives.
- Existing methods for propagating hiPSC-NSPCs have limitations.
Purpose of the Study:
- To develop a novel, convenient protocol for generating and propagating hiPSC-NSPCs.
- To utilize microsphere arrays for efficient neurosphere formation.
- To assess the characteristics and potential of hiPSC-NSPCs generated via this method.
Main Methods:
- Utilized microsphere arrays coated with polyethylene glycol to create non-adhesive surfaces.
- Treated primary human induced pluripotent stem cells (hiPSCs) with noggin to induce neural differentiation.
- Propagated resulting neurospheres as free-floating spheroids.
Main Results:
- Successfully generated neurospheres composed of hiPSC-NSPCs.
- hiPSC-NSPCs exhibited phenotypes nearly identical to hN-NSPCs in marker expression and neuronal differentiation potential.
- Gene expression analysis revealed higher neural and lower glial profiles in hiPSC-NSPCs, with mid-hindbrain regional specificity.
Conclusions:
- The novel microsphere array protocol provides a convenient and efficient method for propagating hiPSC-NSPCs.
- This method supports the generation of neurospheres from hESCs and hiPSCs.
- The findings advance the application of hiPSC-NSPCs in regenerative medicine and neurobiological research.
