Development of microfluidic organ-on-chip systems with rapid temperature control for optical nanoscopy

Project 7 - Matteo Boninsegna

Cherry Biotech

Objectives:

(1) Development of microfluidics organ-on-chip platforms that allow for the maintenance and support of organ functions by perfusing tissue sections while facilitating high speed, high resolution imaging of cellular morphology

(2) Implementation of rapid temperature changes through steering of fluid flow facilitates short-term very high resolution imaging and deceleration of physiological function

(3) Nanoscopy of fenestration dynamics in perfused organ slices

(4) Control of gas partial pressure will enable the control and simulation of physiological stress, such as hypoxia

Expected Results:

(1) Perfusion of liver slices and maintenance of physiological function demonstrated for several hours

(2) rapid fluid exchange with temperature control demonstrated in support of high resolution 3D nanoscopy

(3) Controlling gas partial pressure while maintaining SR-SIM measurements demonstrated

(4) imaging of fenestration dynamics in intact liver sinusoids demonstrated

Liver sinusoidal endothelial cells (LSECs) form particular vessels, called sinusoids, within liver parenchyma and express nanopores on their surface. LSECs also express nanopores on their surface. These nanopores, known as fenestrations, allow the passage of molecules from blood stream to hepatocytes and vice versa. Unfortunately, fenestrations are so small that cannot be observed with common microscopy techniques. Moreover, it is very difficult to culture LSECs in vitro, but also to maintain cut liver sections, without losing their phenotype after few hours. Main aim of DeLIVER project is to shed light on the behavior of fenestrations by implementing high throughput super resolution microscopy (nanoscopy) and organ on a chip technology.

Project Lead:
Dr. Antoni Homs-Corbera

Early Stage Researcher:
Matteo Boninsegna

• Eng Biomedical Engineering - Università degli Studi di Firenze (Italy)
• Single courses (Biomechanics, Bioimaging) - Università degli Studi di Firenze (Italy)
• Internship at the Institute Solid State Physics (ISSP) - University of Latvia
• Leonardo da Vinci grant - Lifelong Learning Program
• Eng Mechanical Engineering - Università degli Studi di Firenze (Italy)
• Sc Biotechnology - Università degli Studi di Firenze (Italy)

I am part of the DeLIVER project as industrial PhD-student. Particularly, I work at CherryBiotech (Rennes, France) which is a microfluidics and organ on a chip startup. Inside Cherry’s R&D team I am developing systems that control different aspects of the cell microenvironment such as temperature, perfusion and much more. Thus, my role is to provide other early stage researchers (ESRs) with devices for LSECs culturing and liver slices maintaining. Comparing to traditional cell culture methods, these platforms will better recapitulate the physiology of liver sinusoids. Furthermore, to carry out fenestration live imaging, the devices will be compatible with ultrafast optical nanoscopes.

Figure 1: One of our last disposable and various control units.

Figure 2: Recent picture of the CherryBiotech team in Lannion (Brittany, France).