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Two-Photon 3D Laser Printing Inside Synthetic Cells.

Tobias Abele1,2, Tobias Messer3, Kevin Jahnke1,2

  • 1Biophysical Engineering Group, Max Planck Institute for Medical Research, Jahnstraße 29, 69120, Heidelberg, Germany.

Advanced Materials (Deerfield Beach, Fla.)
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Summary

Precise 3D hydrogel structures were fabricated inside lipid vesicles using two-photon laser printing. This technique enables controlled positioning of cellular components for bottom-up synthetic biology, including creating functional pores for cargo transport.

Keywords:
3D laser printingPEGDA hydrogeladditive manufacturingbottom-up synthetic biologydirect laser writinggiant unilamellar lipid vesiclestransmembrane pores

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Area of Science:

  • Synthetic biology
  • Biomaterials engineering
  • Microfluidics

Background:

  • Reconstituting cellular components in lipid vesicles is key for bottom-up synthetic cell manufacturing.
  • Deterministic positioning of components within these vesicles remains a significant challenge.

Purpose of the Study:

  • To develop a high-precision method for fabricating 2D and 3D hydrogel architectures inside giant unilamellar lipid vesicles (GUVs).
  • To demonstrate the capability of two-photon 3D laser microprinting for creating functional structures within GUVs without compromising their integrity.

Main Methods:

  • Utilized two-photon 3D laser printing to fabricate hydrogel architectures with precise shapes and positions inside GUVs.
  • Employed a water-soluble photoresist introduced via diffusion into the GUVs.
  • Verified that femtosecond two-photon printing does not disrupt the GUV membrane.

Main Results:

  • Successfully manufactured 2D and 3D hydrogel architectures of arbitrary shapes with high precision within GUVs.
  • Demonstrated the creation of a functional transmembrane pore structure via 3D printing.
  • Showcased the transport of biological cargo, such as DNA, through the printed pore into the synthetic compartment.

Conclusions:

  • Two-photon 3D laser microprinting is a viable and precise technique for building complex hydrogel structures within lipid vesicles.
  • This method offers a powerful new tool for bottom-up synthetic biology, enabling controlled component placement and functionalization.
  • The ability to create functional pores opens avenues for controlled cargo delivery and the development of more sophisticated synthetic cells.