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

A bioreactor is a system able to maintain a cell culture in acontrolled environment, aimed at the simulation of a livingorganism.

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 chamberMixing chamberElectronic circuit and

electro-valve boxPeristaltic pump, PCHeating 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: CO2 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

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 dell’apparato.

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