Research Interests 3D-Cell Culture Group, Gottwald, …Outline 14.10.2015 PD Dr. Eric Gottwald Research Interests of the 3D-Cell Culture Group of Eric Gottwald, Karlsruhe Institute

KIT – University of the State of Baden-Wuerttemberg and

National Research Center of the Helmholtz Association

INSTITUTE FOR BIOLOGICAL INTERFACES, KARLSRUHE INSTITUTE OF TECHNOLOGY, GERMANY

www.kit.edu

Research Interests 3D-Cell Culture Group, Gottwald, KIT, Germany

Authors: E. Gottwald, S. Giselbrecht, P. Wuchter, R. Saffrich, T. Boiselle, S. Kolb, A. Neubauer, L. Schad

Outline

14.10.2015 PD Dr. Eric Gottwald

Research Interests of the 3D-Cell Culture Group of Eric Gottwald, Karlsruhe Institute of Technology, Institute for Biological Interfaces, email: [email protected]

I. Development of microcavity array-systems for 3D-cell culture II. Development of artificial stem cell niches III. Cell Printing IV. Development of organotypic MRI-phantoms for MRI-method development

Introduction

Parameter 2D 3D

Handling + -

Costs + -

Upscaling + +-

Organotypic behaviour - +++

….


Comparison 2D vs. 3D-culture systems

Petri dishes, culture flasks, multiwells

gel systems, hollow fibres, sponges

hollow fibres

microcavity arrays

Introduction

complexity/performance



I The microcavity array-system

p-KITChip f-KITChip r-KITChip

manufacturing process steps

10*106 cells/cm2


I The microcavity array-system

Patented microthermoforming process (SMART-technology) for the generation of thin-walled polymer film-based 3D-culture systems

empty microcavities

300 µm

immunofluorescence in microcavities

stained 3D-aggregates in microcavities


1: screws 2: upper/lower lid 3: microscopy slide 4: microcavity array chip 5: outer gasket 6: inner gasket 7: bioreactor body

I The microcavity array-system: bioreactor assembly


I The microcavity array-system: bioreactor flow schemes

Superfusion


I The microcavity array-system: bioreactor flow schemes

Perfusion


I The microcavity array-system: bioreactor periphery

• closed circulation loop • easy to parallelise • Integration of sensors for O2, CO2, pressure etc. possible

medium reservoir

bioreactor

pump gas from gas mixing station


I The microcavity array-system: bioreactor periphery


I The microcavity array-system: bioreactor periphery and gas mixing station

• Modular periphery • Fibre-optic O2-sensors • Fibre-optic pressure sensors • pH-sensors • Temperature sensors • ...


II Development of artificial stem cell niches: the hematopoietic niche

• Three-dimensionality is likely to play an important role in stem cell maintenance • microcavity array-systems are used for fundamental research with regard to the identification of

crucial factors for stem cell maintenance and directed differentiation

DAPI CD34 CD105

Human HSC/MSC-co-culture experiment

DAPI Β-Catenin CD34+

Hep G2/HSC





plasticity of HSC after 14 days of MSC/HSC co-culture in a bioreactor environment resembles that of freshly isolated HSC from cord blood

Rieke, Gottwald et al., Lab Chip 2008; 8: 2206–2213


II Development of artificial stem cell niches: photoreceptor differentiation


II Development of artificial stem cell niches: photoreceptor differentiation

rho4D2 DAPI

microcavity array reference culture

Rieke, Gottwald et al., Lab Chip 2008; 8: 2206–2213

differentiation efficiency of gerbil retinal stem cells in microcavity arrays is as good as in reference system as judged by photoreceptor protein expression rho4D2


III Cell Printing

unmodified printer converted printer

microcavity array chip printing area


III Cell Printing


III Cell Printing

printed Hep G2 cells stained with DAPI in microcavity arrays


III Cell Printing: printing

printed Hep G2 cells stained with DAPI and CellTracker Green ® in microcavity arrays as a „co-culture“ experiment

III Cell Printing: printing

adressing single microcavities is possible (Hep G2 stained with CellTracker Green ®)

PD Dr. Eric Gottwald

Distribution of contrast agent Gd-DOTA in the medium compartment: inflow

IV Development of organotypic MRI-phantoms for MRI-method development

Gottwald et al., Zeitschr. f. Med. Physik 2013; 23: 102 – 110

cross-section of the bioreactor (MRI-image). Red: microcavity array chip, colors represent different concentrations of contrast agent in perfusion mode. Red arrows: inflow (lower arrow) and outflow (upper arrow) direction with time.

PD Dr. Eric Gottwald

Distribution of contrast agent Gd-DOTA in the medium compartment: inflow





microcavity array filling charasteristics



bioreactor wash-out charasteristics


A. Neubauer et al. ISMRM, 2014, Milano, Italy

23Na and the 1H MRI-image of Hep G2 cells in microcavity arrays


Eric Gottwald group: Hanna Lorig, Ph.D. student Cordula Nies, M.Sc. Food Chemistry Rabea Petermann, M.Sc. Biotechnol. David Thiele, Techn. Elisabeth Jóo, B.Sc. Student Stephanie Kolb, M. Sc. Student Stefan Heikenwälder B. Sc. Student Timo Boiselle, M. Sc. Student Maria Hoshi-Paz, Guest Scientist

contact: Karlsruhe Institute for Technology Institute for Biological Interfaces email: [email protected] Phone: +49(0)721/608-22504

mailto:[email protected]

Documents

Research Interests 3D-Cell Culture Group, Gottwald, …Outline 14.10.2015 PD Dr. Eric Gottwald Research Interests of the 3D-Cell Culture Group of Eric Gottwald, Karlsruhe Institute