A functional artificial cell stabilized by a biomimetic cytoskeletons

This is one of two research projects on bioengineering. KU Leuven is the home institution for this project. To view the Melbourne-based partner project, click here.

Cell-based products are being used with great impact in a wide diversity of both novel and long-established therapeutic applications. Erythrocytes, for example, are used to save thousands of lives every day worldwide. Yet, in low- and middle-income countries, their scarcity and unsafe control are endemic burdens that cost lives. Conversely, the high price tag of CAR-T cell products relegates such therapies to last-resort treatments even in developed countries. Therefore, synthetic strategies (i.e. artificial cells) could make such cell products safer and cheaper. In the paradigmatic case of Erythrocytes, the properties of their cytoskeleton are essential to establish and regulate blood flow, which in turn ensures adequate tissue oxygenation. These properties originate from their cortex, a “nanoscopic chassis” composed of multiple proteins that stabilize and anchors the cellular membrane above it. Attempts to reconstruct a similar cortex within giant unilamellar vesicles (GUVs) have so far delivered poor results with structures that cannot recover the properties of native Erythrocyte cortices.

In this project, we will adapt droplet-based microfluidic strategies for the formation of GUVs to integrate them with a “3D nano-printed” biomimetic cortex. Such structures will be obtained via 2-photon nano-lithographic techniques that have been demonstrated to offer the capacity to pattern nanometric scaffolds with dimensions similar to those of native cellular cortices. We will further characterize the physicochemical and functional properties of these cortex-stabilized GUVs in order to artificially recover specific properties (such as shape, deformability and gas exchange capacity) of native Erythrocytes.

This project aims to achieve two main breakthroughs:

1. Artificially construct a 3D cytoskeletal nano-structure that reproduces the shape, size and mechanical properties of the spectrin cortex of an Erythrocyte
2. Integrate and anchor this artificial cortex within giant unilamellar vesicles (GUV) in order to build, from the bottom-up, an artificial cell that recovers several relevant characteristics of Erythrocytes:
   1. Biconcave shape;
   2. Mechanical properties; and
   3. Capacity for gas exchange.

To achieve these breakthroughs, we will deploy the following work plan:

1. 3D nano-printed biomimetic cortex.
2. Encapsulation of 3D cortices in GUVs.
3. Physicochemical and functional characterization.