Background:
“Organ-on-a-chip” represent a new wave of 3D cell culture models, originated by microfluidic technology, composed by continuously perfused chambers inhabited by living cells arranged in a 3D organization. By these means, the microfluidic device better mimics the microstructure, dynamic mechanical properties and biochemical functionalities of whole living organs (Microfluidic cell culture, Matthias Mehling and Savas Tay). Under physiological conditions, all cell types need a flux of nutrients and waste products: the flow conditions in microfluidic devices mimic this process more closely than in vitro culturing in wells plates. The goal is not to build a whole living organ but rather to synthesize minimal functional units that recapitulate tissue- and organ-level functions. Successful examples of organ-on-a-chip have demonstrated that it is possible to mimic the microarchitecture and functions of living organs, such as lung, heart, and intestine (Creating Living Cellular Machines, R D. Kamm and R Bashir, Annals of Biomedical Engineering), as well as to model diseases (Engineered In Vitro Disease Models, Kambez H. Benam, Donald E. Ingber Annu. Rev. Pathol. Mech. Dis. 2015).
We are part of the Cardiovascular Medicine Unit and our main interest is to study the biological mechanisms underlying thrombosis and aortic aneurisms formation. In collaboration with expert engineers of nanomaterials and microsystems technology in Uppsala University we have designed different microfluidic devices that could mimic the vein vessel and the aortic wall.

In this project we aim to optimize endothelial and smooth muscle cells growth into the chip, test the flow of different cells in suspension that will mimic the blood flow, stain specific markers for every cell type and set up an ideal physiological experiment in which we will investigate disease mechanisms.
The main focus of the project is to set up a “coagulation-on-a-chip” and an “aorta-on-a-chip” model in which we will test the effect of different cytokines, the effect of the silencing of essential genes, and we will study in deep the mechanisms by which different cell populations communicate.

The student will be using the following techniques:
-cell culture
-gene silencing (siRNA)
-cell staining
-Immunofluorescence and confocal microscopy
-RNA extraction and quantification by real-time PCR
-Protein immunodetection by Western blot

Expected Learning Outcomes:
-the student will be able to develop and acquire expertise in a cell culture laboratory, where he or she will work with different cell types (endothelial, smooth muscle, monocytes and lymphocytes cell lines: HUVEC, EAhy, THP-1 and Jurkat respectively)
-The student will understand the concept of gene silencing and will be able to perform gene silencing experiments in vitro using siRNA
-The student will learn different protein immunodetection assays (Immunofluorescence and Western blot) and he or she will be familiar with the confocal microscopy
-The student will get a general knowledge of the basics of cardiovascular disease and more specifically the main mechanisms underlying thrombosis risk and aortic aneurism.

Seminars and Report:
It is expected that the student will attend to the Cardiovascular Medicine weekly seminars and will actively participate in Journal Clubs and scientific meetings as a member of the Unit. An oral presentation will be given to the one of our group meetings to present the results of the project.