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Local Organising Committee
Alessia Di Giancamillo Laura Mangiavini
Laura de Girolamo Antongiulio Marmotti
Celeste Scotti
BRIDGING THE GAP
BETWEEN SURGERY AND BIOLOGY:
HANDS-ON CARTILAGE
LABORATORY COURSE
Honorary President : Matteo Denti, Italy
Program Chair: Giuseppe M. Peretti, Italy
Helder Pereira, Portugal
Congress Chairs: Giuseppe M. Peretti, Italy
EVENT CO-ORGANIZED BY ESSKA
CARTILAGE COMMITTEE AND
RESEARCH COMMITTEE
April 21 - 22 2016
DIBIT
Fondazione Centro San Raffaele del Monte Tabor
Honorary President
Matteo Denti
Program Chair Giuseppe M. Peretti
Congress Chairs
Giuseppe M. Peretti Helder Pereira
Per migliorare la pratica clinica ci serve più ricerca traslazionale! Lo scopo di questo corso è quello di permettere ai chirurghi ortopedici di prendersi una pausa dalla pratica clinica per studiare più approfonditamente la riparazione e rigenerazione cartilaginea. Il programma sarà focalizzato sui diversi aspetti della ricerca in campo cartilagineo, dal prelievo di tessuto alla coltura cellulare e alle più basilari tecniche di biologia molecolare e di istologia. Durante il corso, i partecipanti proveranno una vera esperienza di laboratorio conducendo loro direttamente diversi esperimenti; inoltre, i partecipanti acquisiranno una base teorica sulla riparazione cartilaginea fondamentale sia per intraprendere un eventuale cammino di ricerca di base nel campo della rigenerazione articolare, sia nel comprendere più a fondo i principi che stanno alla base delle moderne metodiche di ingegneria dei tessuti, che al giorno d’oggi sono utilizzati quotidianamente nella pratica clinica. Questo corso rappresenta quindi un’opportunità unica per ricercatori e chirurghi che desiderano avere un approccio diretto alla ricerca in campo cartilagineo.
For a better clinical practice, we need more translational research! The aim of this course is to allow the orthopedic surgeons to “take a short break” from patients practice in order to “go deeper” inside cartilage repair and regeneration. The program will be focused on the different aspects of cartilage research, from tissue harvesting to cell culture and basic techniques of molecular biology and histology. The attendees will experience a real laboratory practice by directly conducting several lab experiments. Additionally, the attendees will acquire a theoretical background on cartilage repair, fundamental both for the research career in joint regeneration and for a greater comprehension of tissue engineering basic principles, which are nowadays routinely applied in clinical practice. Thus, this course represents a unique opportunity for scientists and surgeons, who desire a more direct approach on cartilage research.
Laura de Girolamo (Milan, Italy)
Matteo Denti (Milan, Italy)
Alessia Di Giancamillo (Milan, Italy)
Neil Fisher (Milan, Italy)
Laura Mangiavini (Milan, Italy)
AntonGiulio Marmotti (Turin, Italy)
Silvia Mattia (Turin, Italy)
Carlotta Perucca Orfei (Milan, Italy)
Helder Pereira (Portugal)
Giuseppe M. Peretti (Milan, Italy)
Celeste Scotti (Milan, Italy)
Irene Tessaro (Milan, Italy)
Marco Viganò (Milan, Italy)
FACULTY
TUTORS
Registration Introduction and presentation of the course
THEORY
Articular Cartilage morphology, biochemistry and biomechanics. Growth plate cartilage
The articular environment Stem cells (MSC, ASC and mesenchymal cells
from umbilical cord) Growth factors and Platelet Rich Plasma (PRP) Gene therapy Culture of chondrocytes: from monolayer to
tridimensional (3D) culture
Coffee break
Molecular biology for gene expression analysis: RNA extraction, purification, quantification and measure of RNA integrity
RT PCR PCR; semi-quantitative PCR and real time PCR Fluorocitometric analysis: Fluorescence
Activated Cell Sorter (FACS) Analysis of protein expression and cell
proliferation of chondrocytes both in monolayer and 3D cultures
Use of different cell markers to determine cell phenotype and proliferation
Immunohistochemistry an immunofluorescence of mesenchymal stem cells (differentiated versus undifferentiated)
Immunohistochemistry and immunofluorescence of umbilical cord mesenchymal stem cells
Lunch break
First day (April 21st 2016)
8.30-9 am 9-9:30 am 9:30 am-1 pm
1 - 2 pm
THEORY
Engineered cartilage: animal models. Transgenic mouse models
Biological materials for engineered cartilage Introduction to lab activity
PRACTICE
Articular explants from pig knee, shoulders and elbows
Preparation of articular explants Preparation of the sterile field Collection of articular cartilage Preparation of culture medium, PBS with
antibiotics/antimycotic and collagenase solution Basics on sterility in the use of the laminar flow
cabinet Enzymatic digestion of articular cartilage
First day (April 21st 2016)
2-2:40 pm
2.45 - 6 pm
1 - 2 pm
PRACTICE
Collection of chondrocytes Evaluation of cell number and viability In vitro cell culture: cell seeding and feeding;
passaging, cell counts and viability 3D culture system: use of fibrin glue and
collagen scaffold
Lunch break
THEORY
Histology evaluation score, problems in the biopsy, preservation and storage
Histology and staining on tissue sections: 1. Hematoxylin and Eosin (H&E); different Hematoxylins 2. Safranin-O 3. Alcian Blue 4. Von Kossa 5. Masson’s Trichrome 6. TRAP 7. Immunohistochemistry and immunofluorescence 8. In situ hybridization
PRACTICE
H&E Safranin-O staining and sample analysis under
microscope Biochemistry assays: quantification of DNA and
GAGs in 3D cultures versus native cartilage
End of the Course
Second day (April 22nd 2016)
9am - 1 pm
1 – 2 pm
2 -3 pm
3 - 5 pm
DIBIT Fondazione Centro San Raffaele del Monte Tabor San Raffaele Congress Centre Via Olgettina, 58 - 20132 Milano
Registration
€ 480,00 (taxes included)
Registrations are limited to maximum 20
attendees and will be accepted in the order in
which they are received. Applications will have
to the administrative office along with a copy of
the money tranfer receipt. In case of cancellation,
there will be no refund.
Course fee includes:
Attending to the entire theorical and
practical sessions
Lunch during the I and II course days
Abstract book
Keep International Via Giuseppe Vigoni, 11 - 20122 Milano Tel. +39 0254122579 – Fax +39 0254124871 E.mail: [email protected]
www.keywordeuropa.com
GENERAL INFORMATION
Venue
Registration
Organising
Secretariat
1 - 2 pm