Reversible Intracellular Gelation of MCF10A Cells Enables Programmable Control Over 3D Spheroid Growth

In nature, some organisms survive extreme environments by inducing a biostatic state wherein cellular contents are effectively vitrified. Recently, a synthetic biostatic state in mammalian cells is achieved via intracellular network formation using bio-orthogonal strain-promoted azide-alkyne cycload...

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Bibliographic Details
Published inAdvanced healthcare materials Vol. 13; no. 7; p. e2302528
Main Authors McNally, Delaney L, Macdougall, Laura J, Kirkpatrick, Bruce E, Maduka, Chima V, Hoffman, Timothy E, Fairbanks, Benjamin D, Bowman, Christopher N, Spencer, Sabrina L, Anseth, Kristi S
Format Journal Article
LanguageEnglish
Published Germany 01.03.2024
Subjects
Animals
Cell Survival
Hydrogels - pharmacology
Mammals
Polyethylene Glycols - pharmacology
intracellular crosslinking
biostasis
metabolism
spheroids
hydrogel
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Summary:In nature, some organisms survive extreme environments by inducing a biostatic state wherein cellular contents are effectively vitrified. Recently, a synthetic biostatic state in mammalian cells is achieved via intracellular network formation using bio-orthogonal strain-promoted azide-alkyne cycloaddition (SPAAC) reactions between functionalized poly(ethylene glycol) (PEG) macromers. In this work, the effects of intracellular network formation on a 3D epithelial MCF10A spheroid model are explored. Macromer-transfected cells are encapsulated in Matrigel, and spheroid area is reduced by ≈50% compared to controls. The intracellular hydrogel network increases the quiescent cell population, as indicated by increased p21 expression. Additionally, bioenergetics (ATP/ADP ratio) and functional metabolic rates are reduced. To enable reversibility of the biostasis effect, a photosensitive nitrobenzyl-containing macromer is incorporated into the PEG network, allowing for light-induced degradation. Following light exposure, cell state, and proliferation return to control levels, while SPAAC-treated spheroids without light exposure (i.e., containing intact intracellular networks) remain smaller and less proliferative through this same period. These results demonstrate that photodegradable intracellular hydrogels can induce a reversible slow-growing state in 3D spheroid culture.
ISSN:2192-2659
DOI:10.1002/adhm.202302528