Presentation on theme: "Cells colture Cells culture is an essential tool in biological science, clinical science, and biomedical studies This approach is a fundamental step in."— Presentation transcript:
Cells colture Cells culture is an essential tool in biological science, clinical science, and biomedical studies This approach is a fundamental step in preclinical drug testing There is great interest from pharmaceutical industry to employ it Drug testing involves a large number of tests on similar cells cultures
High Throughput Screening (HTS) HTS, is a methodology for scientific experimentation, especially used in drug discovery It is based on a brute-force approach to collect a large amount of experimental data in less time and with less animals. HTS is achieved nowadays using multi-well equipment, to contain the cell cultures subject to the treatments
Funziona? The multi-well system, however, suffers from a significant problem that may affect the relevance of tests. the environment discrepancy problem comport that the collected data are not directly usable in drug testing. This seems to be a paradox since the multi-well has been the core element of the HTS The environment discrepancy problem lies in the fact that the tissue grown in wells is only a brutal approximation of the biological reality.
Our Idea Make a bioreactor system a HTS machine Autonomus Sicuro Usefull Easy to use A bioreactor is a system able to maintain a cell culture in a controlled environment, aimed at the simulation of a living organism.
The Bioreactors Bioreactors have not been designed for HTS We present a new bioreactor, called MCB, able to perform high throughput experiment in a in vivo-like simulated environment for a long time Bioreactor is a better approximation of in vivo physiology.
The MCB System The bioreactor system consists of the following parts: Cell culture chamber Mixing chamber Electronic circuit and electro- valve box Peristaltic pump, PC Heating system
Cells colture chambers This is the core of the bioreactor system and contains the cell mono-layer where cells are treated. It is made of PDMS Poly- dimethyl-siloxane, a biocompatible silicone polymer. We have developed several types of cell culture chambers, every one can be plugged into the system
The mixing chamber The mixing chamber serves for pH and oxygen regulation. In the chamber the medium is perfused with gas according to the measured pH pH regulation is performed by inserting two different gases in the mixing chamber: CO 2 and Air
The Control System The micro controlled board runs the μTNetOS operating system The goal of the application is to make the bioreactor autonomous by monitoring the environment variables Ethernet is the communication bus A High Throughput bioreactor experiment, employs many bioreactors with different cell culture chambers. A pathology can be simulated in one of the Bioreactors, and the others are used for a control reference.
µTNetOS μTNetOS is a generated operating system: the system generator takes as input the description of an XML based protocol, The system uses cooperative multi-tasking to run concurrent activities μTNetOS is based on a robotics programming framework called Robotics4.NET The framework proposes a software architecture inspired to the architecture of the human nervous system. The Brain: The Bodymap The Roblets
Robotics4.Net and the Bioreactors Bioreactors µTNetOS [Roblet] PC [Brain and bodymap]
How control Ethernet is the communication bus The graphical application, used to control an experiment, can receive data from several units running in parallel. The network allows connections through Internet, allowing remote monitoring of experiments, A pathology can be simulated in one of the Bioreactors, and the others for a control reference
The High Throughput Bioeactor To control the pH in the medium we developed a dedicated control adaptive algorithm, based on a step strategy. The pH response to the diffusion of CO2 and air is very difficult to predict, because it strongly depend on the environmental variables we do not use the mathematical model of the CO2 and air diffusion in water medium, but we use a function defined by an algorithm. Our system continuously inserts air in
pH control algorithm To control the pH in the medium we developed a dedicated control adaptive algorithm, based on a step strategy. The pH response to the diffusion of CO2 and air is very difficult to predict, because it strongly depend on the environmental variables We do not use the mathematical model of the CO2 and air diffusion in water medium
The user interface The Graphical User Interface is developed in C#.Net Is based on a multi tab structure GUI is used to read data from the bioreactors and to setup the experimental variables There are tools for sensor Calibration Manual control is possible
Results 48h parallel experiment with 4 bioreactors
Sviluppi futuri Il sistema ha dimostrato di funzionare correttamente Le modifiche apportate hanno incrementato notevolmenete la maneggevolezza dellapparato. Possibilità di effettuare esperimenti High Throughput in ambiente simulato Riduzione del numero di cavie necessarie. Prodotto pronto per la commercializzazione Il sistema è pronto per eventuali modifche Inserimento sistemi di allarme via mail o SMS Ulteriori test per definizione costanti di controllo Eliminazione bagnetto termostatico
Results endothelial cells on a laminar flow colture chamber after 48h