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The role of leukocyte-derived microparticles in airway inflammation
Universita degli Studi di Pisa
Dipartimento di Patologia Chirurgica, Medica, Molecolare e di Area Critica
Prof. Alessandro Celi
Dott. Tommaso Neri
Type of Research Project
- Basic science
What is the background of the project?
It has been known for several years that eukaryotic cells are capable of shedding membrane fragments, commonly referred to as ectosomes or microparticles (MP). Originally considered in vitro artifacts devoid of physiological significance, MP are rapidly gaining attention as modulators in numerous processes, including, among others, inflammation and blood coagulation. It has been demonstrated, for example, that MP carry integral membrane proteins like the initiator of blood coagulation, tissue factor, and the cell-cell adhesion molecule ICAM-1. Leukocyte derived MP are capable of promoting inflammation by activating lung epithelial cells. One key question that is currently being investigated concerns the mechanisms of MP generation.
What is the aim of the project?
The aim of the project is to study the mechanisms of MP shedding. Specifically, the role of the ion channel, TRPC6, is a candidate molecule that might represent a critical stimulus for MP generation.
What techniques and methods are used?
- In vitro generation of MP upon stimulation with different agonists - Quantification of MP concentration by Zymuphen MP-activity kit - Measurement of intracellular calcium by Molecular Probes Fluo-4 NW Calcium Assay kit - Measurement of other protein possibly involved in MP generation by commercials kit or by western blot analysis
What is the role of the student?
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student will carry out her/his project according to the tutor’s instructions. The student will first become confident with the basic laboratory procedures: use of cell cultures under sterile conditions, growth of cultured cells, cell counting, preparation of cells for experiments. The student will then be able to begin the experimental part that will consist in stimulating the cells to generate microparticles, enumerating them with different techniques and studying some of the mechanisms underlying their generation.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
During the first days, the tutor will provide materials to the student, for example, papers on the topic of the project and methodological protocols. Moreover tutor will teach the theoretical bases of lab work.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a scientific report - No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
- Enthusiasm for learning - Curiosity - Critical thinking and reasoning - Initiative
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months)
- - Cell-derived microparticles and the lung. Nieri D; Neri T; Petrini S; Vagaggini B; Paggiaro P; Celi A. Eur Respir Rev. 2016 Sep;25(141):266-77. doi: 10.1183/16000617.0009-2016. Review.
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