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Cell spheroid creation by transcytotic intercellular gelation.

Jiaqi Guo1, Fengbin Wang2,3,4, Yimeng Huang1

  • 1Department of Chemistry, Brandeis University, Waltham, MA, USA.

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|May 22, 2023
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Summary

Researchers developed a new method using enzyme-responsive D-peptides to create cell spheroids, which are crucial for regenerative medicine. This process involves peptide transcytosis and gel formation, mimicking natural tissue environments.

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Area of Science:

  • Biomaterials Science
  • Tissue Engineering
  • Regenerative Medicine

Background:

  • Cell spheroids are vital for bridging in vitro and in vivo research, but their induction via nanomaterials is inefficient and poorly understood.
  • Developing efficient methods for cell spheroid formation is critical for advancing regenerative medicine and tissue engineering applications.

Purpose of the Study:

  • To investigate the atomic structure of helical nanofibres self-assembled from enzyme-responsive D-peptides.
  • To elucidate the mechanism by which D-peptides induce cell spheroid formation through transcytosis and intercellular gel formation.
  • To explore the potential of this D-peptide system as a novel approach for regenerative medicine.

Main Methods:

  • Cryogenic electron microscopy (cryo-EM) was used to determine the atomic structure of self-assembled helical nanofibres.
  • Fluorescent imaging tracked the transcytosis of D-peptides and their subsequent interactions within the cellular environment.
  • Enzyme-responsive D-peptides, specifically D-phosphopeptides, were synthesized and utilized for their unique properties.

Main Results:

  • Helical nanofibres were formed from enzyme-responsive D-peptides through endocytosis and dephosphorylation.
  • Secreted nanofibres formed intercellular gels on the cell surface, acting as artificial matrices.
  • These nanofibres facilitated fibronectin fibrillogenesis, leading to the induction of cell spheroids.
  • Cell spheroid formation was dependent on endo-/exocytosis, phosphate triggers, and peptide assembly shape-switching.

Conclusions:

  • This study demonstrates a novel mechanism for inducing cell spheroid formation by coupling peptide transcytosis with morphological transformation.
  • The developed D-peptide system shows significant potential as a biomaterial for regenerative medicine and tissue engineering.
  • Understanding the interaction between D-peptide assemblies and extracellular matrix components like fibronectin is key to optimizing spheroid formation.