Biophysical Engineering Group, Max Planck Institute for Medical Research, Jahnstraße 29, 69120, Heidelberg, Germany.
Department of Physics and Astronomy, Heidelberg University, 69120, Heidelberg, Germany.
Adv Mater. 2022 Feb;34(6):e2106709. doi: 10.1002/adma.202106709. Epub 2021 Dec 26.
Toward the ambitious goal of manufacturing synthetic cells from the bottom up, various cellular components have already been reconstituted inside lipid vesicles. However, the deterministic positioning of these components inside the compartment has remained elusive. Here, by using two-photon 3D laser printing, 2D and 3D hydrogel architectures are manufactured with high precision and nearly arbitrary shape inside preformed giant unilamellar lipid vesicles (GUVs). The required water-soluble photoresist is brought into the GUVs by diffusion in a single mixing step. Crucially, femtosecond two-photon printing inside the compartment does not destroy the GUVs. Beyond this proof-of-principle demonstration, early functional architectures are realized. In particular, a transmembrane structure acting as a pore is 3D printed, thereby allowing for the transport of biological cargo, including DNA, into the synthetic compartment. These experiments show that two-photon 3D laser microprinting can be an important addition to the existing toolbox of synthetic biology.
为了从底层制造合成细胞这一宏伟目标，各种细胞成分已经在脂质体内部被重新组装。然而，这些成分在隔室内部的确定性定位仍然难以捉摸。在这里，通过使用双光子 3D 激光打印，可以在预先形成的巨大单层脂质体（GUV）内部高精度地制造具有几乎任意形状的 2D 和 3D 水凝胶结构。所需的水溶性光致抗蚀剂通过在单个混合步骤中的扩散进入 GUV。至关重要的是，腔内的飞秒双光子打印不会破坏 GUV。除了这个原理验证演示之外，还实现了早期的功能结构。特别是，作为孔的跨膜结构被 3D 打印，从而允许包括 DNA 在内的生物货物进入合成隔室。这些实验表明，双光子 3D 激光微打印可以成为合成生物学现有工具包的重要补充。
