Cellbasedautonomous

biosensingmicrosystem
LiveSense

Autonomous cell-based biosensor microsystem

A week of autonomy (?)
Communication unit (to GSM device)
Versatile for various cell models and secondary sensors

data com

sampling

Cells
nutrient
supply

bioreactor

optical
sensors
electrical
sensors

signal
processing
unit

comm.
unit

power supply

microsystem platform

Cell-based biosensors provide a biologically relevant

response to toxic compounds and mixtures

The8teamsinLiveSenseproject

EPFL-LMIS

Microsystems Laboratory

Ph. Renaud

Microfluidics, cell chips, bio-impedance

UNIL-DMF

Department of Fundamental Biology

J. van der Meer

Cell biology, gene reporters, bacterial sensors

HESSO-ISI

Industrial Systems Institute

M. Geiser

Microsystems, electronics, optical sensors

ETHZ-MAT

Biologically Oriented Materials

V. Vogel

Biomaterial, cell biology

CSEM

Nanobiotechnology group

M. Liley

Surface biochemistry, biomaterials

EPFL-LEPA

Electrochemistry and Analytics Lab.

H. Girault

Electrochemical sensing, analytical chemistry

EPFL/NE-IMT

UNIL- IST

Sensors and Actuators Laboratory

N. de Rooij

Microfluidics, microsensors

/ P. van der Val

Institute of Occupational Health

M. Riedicker

Health effect of pollution

SECONDARY SENSORS

CELL MODELS

- Electrochemical microsensors
- Fluorescence/biolumin.
Fluorescence/biolumin. sensors
- Impedance sensing
- Sensors validation

- Specification of cell models

- Culture protocols
- Assessment of cell models
- Culture in microbioreactors

MICRO-BIOREACTOR
- Bacterial microbioreactor
- Biomaterials and 3D culture
- Microfluidic platform
- Nutrient and sample supply
- Integration of monitoring sensors

SYSTEM INTEGRATION
- Systems specifications
- Electronic platform + demo
- Bioreactor integration
- Validation of bioreactor control
- Demonstration

Cellmodels
Gene=callymodiedbacteria(Ecoli)
Fluorescenceorelectrochemicaldetec=on

Epithelialcellslines:formingepithelium=ssues,oneor
severallayers,formscelljunc=ons
TransEpithelialElectricalResistance(TEER)

Fibroblastscelllines:structuralframeworkforanimal
=ssues
Mechanoop=calbronec=nsensing

Hepatocytes(primaryofcelllines):7080%oftheliver's
cytoplasmicmass
Fluorescence,electrochemicalorimpedancedetec=on

LMIS

Secondarysensors
Bioreactormonitoring
LEPA

Measuresalinitybyconduc=vity
MeasureDissolvedOxygen
MeasurepH
Measureglucoseandlactate
+otherenzyma=csensors
Cellmetabolismmonitoring
LMIS

Measurecellimpedance
MeasureTransEpithelialresistance
Measureuorescence

Bacterialcells:E.coliinbeads

Detec=onsystemisbasedontheinternalregula=onmechanismofbacteria
againstchemicalstress
AGFPreportergeneisassociatedtoaregulatorygeneinordertoproduceadirect
uorescencesecondarysignal

Chemical target:
2-hydroxybiphenyl
Reporter protein: gfp
Induction time: 4 h

Bacterialcellsinmicrouidics

Microuidictrapsforbacteria
Con=nuousowinduc=on
Insituuorescencedetec=on
Arsenicdetec=oninwater

EpithelialcellsandTEER
RTGill-W1 cell line (fish)
Cultured between 18-21C
Grow under atmospheric CO2 conditions
Cell line did NOT form tight junctions
Caco-2 cells (intestestine cell line)
Known high TEER values
Looking for primary fish cell lines that form tight junctions

Measure Trans Epithelial

Electrical Resistance (TEER)
of epithelial cells on ultra-thin
porous silicon nitride
membranes.

Electrochemicalsecondarysensors

Microuidicamperometricenzyma=cglucose+lactatesensor
Indirectmonitoringofcellmetabolism
Verysmallsamplevolume.Insitumeasurement
Deposition Points
Sample
Inlet

PBS Inlet

Outlet

Electrochemicalsecondarysensors
Mul=sensorpla[orm
Placedupstreamanddownstream
frombioreactor

LEPA

Electrochemicalsecondarysensors
Glucoseoxidaseonmagne=cbeads
Directdownstreamdetec=onofH2O2onsilver

Channel height

Interelectrode gap
=electrode width

LEPA

Bioreactorforhepatocytes

HepG2cellline:knownmodelfortoxicology
Alsopossiblewithprimaryhumancells

LMIS

Hepatocytes:impedancemonitoring
Monitoringofcellgrowthinculture
Exposuretotoxicants=changeofimpedance

Cell growth

0.1mM SDS toxicity

LMIS

50
0

af
te
r2
.5
h

D
ay
1
D
ay
2
D
ay
3
D
ay
4
D
ay
5
D
ay
6
D
ay
7

50

100

af
te
r1
.5
h

100

150

af
te
r1
5m
in

150

200

5m
in

200

250

D
ay
7
af
te
r

Relative impedance / %

M Relative impedance / %
ed
iu
m

250

Mechanoop=calbiomolecularsensing
(basedonuorescenceresonanceenergytransfer)

D: Alexa 488
A: Alexa 546

donors random on lys, acceptors on cys

N

D A

DA
C

SS
SS

DA

Biochemical labeling of plasma Fibronectin,

added to cell medium,
harvested by cells,
incorporated into extra-cellular matrix

Baneyx, Baugh,Vogel, PNAS 98 (2001) 14464 & PNAS 99 (2002) 5139.

Smith, Gourdon, Kubow, Little, Vogel, PLOS Biology 5 (2007) e268

Mechanoop=calsensing+broblasts

Celldeathwillreduce
extracellularmatrix
tension

Smith, Gourdon, Kubow, Little, Vogel, PLOS Biology, 5 (2007) e268

Little, Smith, Ebneter, Vogel, Matrix Biol. 27 (2008) 451

Systemintegra=on

data com

sampling

Cells
nutrient
supply

bioreactor

optical
sensors
electrical
sensors

signal
processing
unit

comm.
unit

power supply

microsystem platform

Systemintegra=on:bioreactor+sensors

Autonomoussensorpla[orm

Refractometrysensor(sugarinwinegrapes)
BluetoothcomtoGSMdevice

Nextsteps
SECONDARY SENSORS
CELL MODELS
- Specification of cell models
- Culture protocols
- Assessment of cell models
- Culture in microbioreactors

MICRO-BIOREACTOR
- Bacterial microbioreactor
- Biomaterials and 3D culture
- Microfluidic platform
- Nutrient and sample supply
- Integration of monitoring sensors

- Electrochemical microsensors
- Fluorescence/biolumin.
Fluorescence/biolumin. sensors
- Impedance sensing
- Sensors validation

SYSTEM INTEGRATION
- Systems specifications
- Electronic platform + demo
- Bioreactor integration
- Validation of bioreactor control
- Demonstration

LiveSenseindustrysupport

Lonza

Cell culture products

access to cells and culture kits

Ayanda Biosystems

Bioelectric platform

platform, microelectrodes

Debiotech

Drug delivery devices

micro-pump samples

Novozymes

Protein engineering, HTS

access to cell based assays

Neroxis/Veolia

Chemical microsensors

sensor samples

Diagnoswiss

Electrochemical immunosensing

expertise, sensor samples

Biocartis

Multiplex diagnostics

protein microcarriers

Trade-4

Telemedecine

technical support

European centre for validation of alternative methods

expertise, standards