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: +49 711 970-4152sibylle.thude@igb.fraunhofer.de

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Thanks for your attention!!Thanks for your attention!!

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