Test systems at Fraunhofer IGB systems at Fraunhofer IGB Sibylle Thude ... collagene matrix with...
Transcript of Test systems at Fraunhofer IGB systems at Fraunhofer IGB Sibylle Thude ... collagene matrix with...
Test systems at Fraunhofer IGBTest systems at Fraunhofer IGB
Sibylle ThudeSibylle Thude
Workshop: Micro-fluidics and nano-biotechnology for the health care industry, IPT, São Paulo, Brazil, 23rd March 2011
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Overview
Tissue Engineeringg g
Avascular Test Systems - skin
di d ( ki )- accredited test systems (skin / Caco-2)
Three-dimensional, vascularized matrix „BioVaSc“
Bioreactors
Vascularized Test Sytems - Trachea
- Small intestine
- Liver
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Department of Cell and Tissue Engineering
Avascular Test SystemsAvascular Test Systems
Vascularized Test Systems
Cells and Biomaterials
Bioreactors for Tissue Engineering
Toxicology and Accreditationgy
Attract GroupCardiovascular TissueCardiovascular Tissue
GMP Production of cell-based products
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Tissue Engineering
C tII.
Concept
I I l ti f i ll
I.
I. Isolation of primary cells
II. Culture and proliferation in vitro
III. Growth on 3D scaffoldV
IV. Human in vitro test tissue
V Implantation as transplantIII.
V.
V. Implantation as transplant
test system IV.
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Tissue Engineeringg g
Supporting scaffold
structured surface - micro- / nano-structured
- lines / globular structures
functionalization - chemicalfunctionalization chemical
- organic modification
- biochemical
biocompatibel / biological - BioVaSc
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Avascular Test systems – 3D Skin Model
Biocompatibility test according toDIN EN ISO 10993 5 DIN EN ISO 10993-5
irritation potential of cosmetica(REACH)(REACH)
penetration / permeation studies
Investigation of
- metabolism
- apoptosis / necrosis
- inflammation
pathologic models
- wound healing
- infection
- aging
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Accredited test systems
Accredited material testing according to DIN ISO 10993-5 (Biocompatibility/Cytotoxicity)10993-5 (Biocompatibility/Cytotoxicity)
on monolayer cultures using different cell lines (H C T MRC 5 / L929) Primary human (HaCaT, MRC-5 / L929)
employing a three-dimensional, organoid skin model
Primary human keratinocytes
model
on extracted material or in direct contact
3D skin model
Accredited Caco-2 model for transport studiesAccredited Caco 2 model for transport studies
Classification of substances with regard to their intestinal solubility and permeabilitytheir intestinal solubility and permeability
Epithelial cell line (Caco-2)
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Three-dimensional, vascularized matrix
BioVaSc (Biological Vascularized Scaffold)g
Segment of porcine Jejunum g p j
collagene I and III
cell free
capillary system remains p y y
reseeding with human cells
connection to capillary systemis possible
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Three-dimensional, vascularized matrix
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Three-dimensional, vascularized matrix
vascular structure
matrix
arterial influxVenous efflux arterial influxVenous efflux
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Three-dimensional, vascularized matrix
Functionality is proven by implantation
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Bioreactors
Development of Bioreactors
Simulation of in vivo situation
- blood pressurep
- dynamic cultivation (shear stress on cells)
- airlift cultivation (e.g. trachea)( g )
different bioreactors
i ifi l i i- tissue specific cultivation
- trachea / small intestine / liver
Computer aided monitoring
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Bioreactors – in vivo simulation
Simulation in BioreactorIn vivo situation
chemical interaction
mechanical
ti l tistimulation
cell-cell-interaction
cell-matrix-interaction
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Bioreactors – for different test systems
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Bioreactors - Computer aided monitoringg
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Vascularized Test Sytems based on BioVaSc matrixy
Trachea Test System Trachea Test System
Simulation of penetration / resorption via respiratory tract
nano-encapsulated drug application via aerosol
Small Intestine Test System
Resorption via intestinale barrierResorption via intestinale barrier
Investigation of new formulations
Liver Test System
Development of new drugs / therapies
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Trachea Test System
I l i f i h h l Isolation of primary human tracheal epithelial cells from donor material
Generation of tissue cultures on collagen scaffolds
Chemical and mechanical Chemical and mechanical stimulation to initiate cilia formation in the bioreactor
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Trachea Test Systemy
Air flow
medium flowflow
li iApplication areas
Testing of new (stent-) materialsg
Testing of chemicals or active substances for penetration behaviour
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Small Intestine Test System
two compartments
- apical = cell line Caco-2
- basal = endothelial cells
in vitro simulation of gut-blood-barrier
b ti / ti t tabsorptive / secretive transport
high prismatic cell proliferation
cell barrier sustained despite shear stress
ll t i ith ll
increased paracellular permeability
collagene matrix with cells
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Liver Test System
Benefits compared to 2D model
prolongated cultivation of hepatocytes without loss of liver specific prolongated cultivation of hepatocytes without loss of liver specific activities
Synthesis of urea, albumine and lactateSynthesis of urea, albumine and lactate
Phase I and II metabolism
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GMP production of cell-based products
l l A D di
quality control
cleanrooms class A–D according to EU-GMP-Guideline
class Dcontrol
separate locks for personnel and material
production cleanrooms
2 QC laboratories - class D
d i lPlanned for expansion 4 production rooms - class B / C
material room and cold storage -
class Cclass B
material room and cold storage class D
Total area 216 m2
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GMP production of cell-based products
Manufacturing authorization granted
Autologous bone tissue based on adult stem cells (Aastrom Biosciences)
Autologous blood vessel implant (Vaso Tissue)
Autologous cartilage transplant
melanocyte transplant
Applied
Autologous melanocyte transplantHMB45 vimentin
y pAutologous melanocyte transplant
Vascularized transplant for trachea and esophagus surgery (clinical study and esophagus surgery (clinical study for European ATMP authorization)
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Interactions with Micro-fluidic and Nano-Biotechnology
Micro-fluidic Technology
Development of high-delivery micro-pumps for different bioreactors
- controlled flux of cell medium during cultivation
- physiological pressure / fluidic profile
- improved proliferation of cellsp p
- high throughput testing
Nano BiotechnologyNano-Biotechnology
Development of new formulations and their investigation on
Ski T t S t ( l / l i d)- Skin Test System (avascular / vascularized)
- Small Intestine Test System
T h T S- Trachea Test System
- Liver Test System
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Attract Group Cardiovascular Tissue Engineering
Tissue Engineering of an “optimal” heart valve:
To identify the phenotype of semilunar valves cells in human heart valves (pulmonary valve and aortic valve)
Identification and characterization of extracellular Identification and characterization of extracellular matrix proteins in developing heart valves
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Attract Group Cardiovascular Tissue Engineering
Tissue Engineering of an “optimal” heart valve:
To identify suitable autologous progenitor cells
To design optimal scaffold substrates (electrospinning)
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Your partner from Fraunhofer IGB
Sibylle ThudeGroup manager „Accreditation and Toxicology“Fon: +49 711 [email protected]
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Thanks for your attention!!Thanks for your attention!!
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