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Transfecting and Nucleofecting Human Induced Pluripotent Stem Cells
Published on: October 5, 2011
Electrically guiding migration of human induced pluripotent stem cells
Jiaping Zhang1, Marco Calafiore, Qunli Zeng
1Department of Dermatology and Department of Ophthalmology, Institute for Regenerative Cures, UC Davis School of Medicine, Davis, CA 95817, USA.
Stem Cell Reviews and Reports
|March 5, 2011
Summary
Physiological electric fields (EFs) effectively guide human induced pluripotent stem (hiPS) cells, a promising alternative to embryonic stem (ES) cells, in both 2D and 3D environments for improved cell therapy applications.
Area of Science:
- Regenerative Medicine
- Biophysics
- Cell Biology
Background:
- Stem cell therapy faces challenges with poor cell homing and integration into host tissues.
- Human induced pluripotent stem (hiPS) cells offer a viable alternative to embryonic stem (ES) cells for therapeutic applications.
- Effective guidance of transplanted stem cells is crucial for successful cell therapy.
Purpose of the Study:
- To investigate the feasibility of using physiological electric fields (EFs) to guide the migration of hiPS cells.
- To determine the response of hiPS cells to applied EFs in both two-dimensional (2D) and three-dimensional (3D) culture systems.
- To compare the EF-directed migration of hiPS cells with that of hES cells.
Main Methods:
- Applied small, physiological electric fields (<30 mV/mm) to cultured hiPS cells in 2D and 3D environments.
- Assessed cell migration directionality and sensitivity in response to EFs.
- Analyzed the expression of pluripotency markers (SSEA-4, Oct-4) after EF exposure.
- Compared galvanotaxis of hiPS cells and human embryonic stem (hES) cells.
- Investigated the effect of Rho-kinase (ROCK) inhibition on EF-directed migration.
Main Results:
- Applied EFs stimulated and guided hiPS cell migration towards the anode in a concentration-dependent manner.
- hiPS cells exhibited directional migration in 3D cultures under EF stimulation, unlike stationary cells in control conditions.
- EF exposure did not affect the expression of pluripotency markers SSEA-4 and Oct-4.
- hiPS cells demonstrated greater sensitivity and directedness to EFs compared to hES cells, which migrated towards the cathode.
- ROCK inhibition increased hiPS cell motility but significantly reduced their directionality in EFs (70-80%).
Conclusions:
- Physiological electric fields are effective guidance cues for hiPS cell migration in both 2D and 3D environments.
- EF-guided migration of hiPS cells is dependent on Rho-kinase (ROCK) signaling pathways.
- These findings suggest potential in vivo applications of EFs for directing transplanted stem cell migration in regenerative medicine.
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