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Reversible Intracellular Gelation of MCF10A Cells Enables Programmable Control Over 3D Spheroid Growth.
Delaney L McNally1, Laura J Macdougall1,2, Bruce E Kirkpatrick1,2,3
1Department of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, CO, 80303, USA.
Advanced Healthcare Materials
|December 24, 2023
Summary
Scientists created a reversible biostatic state in mammalian cells using intracellular hydrogels. This synthetic biostasis slows cell growth and metabolism, offering potential for controlled cell proliferation in 3D cultures.
Area of Science:
- Biotechnology
- Cell Biology
- Materials Science
Background:
- Organisms naturally enter biostasis (a vitrified state) to survive extreme conditions.
- Synthetic biostasis in mammalian cells is achieved using intracellular networks formed by bio-orthogonal strain-promoted azide-alkyne cycloaddition (SPAAC) reactions.
- Functionalized poly(ethylene glycol) (PEG) macromers facilitate intracellular network formation.
Purpose of the Study:
- To investigate the effects of intracellular network formation on a 3D epithelial MCF10A spheroid model.
- To explore the induction of a reversible biostatic state in mammalian cells.
- To assess the impact of intracellular hydrogels on cell proliferation and metabolism.
Main Methods:
- MCF10A cells were transfected with macromers and encapsulated in Matrigel to form 3D spheroids.
- Strain-promoted azide-alkyne cycloaddition (SPAAC) reactions were used to form intracellular hydrogel networks.
- Photosensitive nitrobenzyl-containing macromers were incorporated for light-induced network degradation.
Main Results:
- Intracellular hydrogel network formation reduced spheroid area by approximately 50%.
- The hydrogel network increased the quiescent cell population (indicated by p21 expression) and reduced bioenergetics (ATP/ADP ratio) and metabolic rates.
- Light-induced degradation of the PEG network reversed the biostasis effect, restoring cell state and proliferation.
Conclusions:
- Photodegradable intracellular hydrogels can successfully induce a reversible slow-growing state in 3D spheroid cultures.
- This approach offers a novel method for controlling cell proliferation and metabolic activity.
- The findings have implications for regenerative medicine, tissue engineering, and drug discovery.
