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Stencil Micropatterning of Human Pluripotent Stem Cells for Probing Spatial Organization of Differentiation Fates
Published on: June 17, 2016
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Patterned substrates fabricated by a controlled freezing approach and biocompatibility evaluation by stem cells
Lei Qian1, Adham Ahmed1, Laurence Glennon-Alty2
1Department of Chemistry, University of Liverpool, Oxford Street, Liverpool L69 7ZD, UK.
Summary
Controlled freezing fabricates aligned cell culture patterns on various materials. PLGA-chitosan and silica substrates effectively guided mouse mesenchymal stem cell alignment, offering stable, biocompatible surfaces for long-term cell culture.
Area of Science:
- Biomaterials Science
- Cell Biology
- Surface Engineering
Background:
- Patterned substrates are crucial for controlling cell growth and alignment.
- Existing fabrication techniques include photolithography, soft lithography, and microcontact printing.
- A facile and tunable method for creating biocompatible surface patterns is needed.
Purpose of the Study:
- To develop a controlled freezing approach for fabricating aligned and grid surface patterns.
- To investigate the biocompatibility and cell alignment capabilities of these fabricated substrates.
- To evaluate the long-term stability of different patterned materials for cell culture.
Main Methods:
- Fabrication of aligned and grid surface patterns using a controlled freezing technique.
- Demonstration of pattern fabrication with polymers, nanoparticles (organic/inorganic), and mixtures.
- Evaluation of substrate biocompatibility using WST-8 viability tests (cell counting kit-8) and culturing of mouse mesenchymal stem cells (mMSCs).
Main Results:
- Aligned surface pattern spacing was tunable by adjusting freezing rates.
- Mouse mesenchymal stem cells aligned with the ridges on PLGA-chitosan and silica colloid substrates.
- PLGA substrates showed degradation, limiting their long-term suitability, while PLGA-chitosan and silica substrates remained stable and maintained cell alignment.
Conclusions:
- Controlled freezing offers a versatile method for fabricating tunable surface patterns for cell culture.
- PLGA-chitosan and silica colloid substrates demonstrate excellent biocompatibility and long-term stability for guiding mMSC alignment.
- These stable patterned substrates show promise for applications in tissue engineering and regenerative medicine.
