development of a matrix-based technology platform for the

1
Business Potential New market entry from microbial library screening (industrial biotechnology) to mammalian library screening (pharmaceutical industry) High throughput compatible matrix-based 3D cell culture technology Primary cell line development o Patient-derived cancer (stem) cell lines Phenotypic high throughput screening o Enrichment of cancer-derived cell subpopulations for targeted therapy o Discovery of novel biologics and efficacy and toxicology studies with small molecules Acknowledgements We gratefully thank the innovation promotion agency (CTI, Switzerland) for finacial support of the project and technical guidance Development of a matrix-based technology platform for the high throughput analysis of 3D cell cultures Project Number: CTI 18364.2 PFLS-LS Main Applicant: ZHAW, Dr. Markus Rimann, Dr. Epifania Bono, Armin Picenoni Research Partner: University Hospital Zurich, Dr. Emanuela Felley-Bosco Main Industrial Partner: FGen GmbH, Dr. Andreas Meyer, René Pellaux, Corinna Hund Start: 01.01.2017, Duration: 24 months Project goal The screening of large cell libraries is an important process in pharmaceutical discovery and R&D, e.g. to define drug targets or develop effective medicines. The goal of this project is the implementation of a screening platform based on 3D cultivation of primary human mesothelioma cells encapsulated in alginate hydrogels (Fig. 1) . To this end new hydrogel compositions will be designed, tested and finally utilized in the Nanoliter Reactor (NLR) cultivation system (Fig. 2) that enables high throughput analysis of 3D cell cultures. Key Findings Successful manual cell encapsulation of mesothelioma cell line ZL55SPT into alginate beads (Fig. 1A) Life/dead staining of encapsulated ZL55SPT cells (Fig. 1B) Scientific Innovation 3D cell culture of mammalian cells in Ca-alginate hydrogels offering different microenvironments: o Blending of alginate with natural environment mimicking compounds o Modification of the hydrogel stiffness to adapt to native tissue Implementation of the modified Ca-alginate hydrogel in the NLR-technology to produce reliable monodisperse capsules at high reproducibility. Development of a novel method to generate patient-derived primary mesothelioma cell lines in 3D (e.g. cell subpopulations, like cancer stem cells) from a biopsy in a high throughput manner using the NLR-technology. Retain the encapsulated mesothelioma cells in their native state to use them for drug development/treatment to gain predictive drug responses. Comparison of in vitro to in vivo and come up with strategies to develop personalized drug treatment regimes. Fig. 1: Mesothelioma cell line ZL55SPT encapsulated in alginate beads. A) Cell growth and spheroid formation after 9 days of incubation. B) Life/Dead staining (green/red) of cells after 21 days of incubation. Fig. 2: NLR technology scheme Cultivation 10 6 NLRs/100 mL Encapsulation 0.7 - 3.5 kHz Cell library 10 5 – 10 7 cells Ø: 20 – 600 μm NLRs with cells Phenotypic Assay NLRs with clusters >200 cells/cluster Fluorescent hits 3 fluorescent signals Characterization Secondary assays Analysis/Selection 10 – 250 Hz Isolation from NLR A B brought to you by CORE View metadata, citation and similar papers at core.ac.uk provided by ZHAW digitalcollection

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Page 1: Development of a matrix-based technology platform for the

Business Potential• New market entry from microbial library screening

(industrial biotechnology) to mammalian library

screening (pharmaceutical industry)

• High throughput compatible matrix-based 3D cell

culture technology

• Primary cell line development

o Patient-derived cancer (stem) cell lines

• Phenotypic high throughput screening

o Enrichment of cancer-derived cell subpopulations

for targeted therapy

o Discovery of novel biologics and efficacy and

toxicology studies with small molecules

AcknowledgementsWe gratefully thank the innovation promotion agency (CTI,

Switzerland) for finacial support of the project and technical guidance

Development of a matrix-based technology platform

for the high throughput analysis of 3D cell culturesProject Number: CTI 18364.2 PFLS-LS

Main Applicant: ZHAW, Dr. Markus Rimann, Dr. Epifania Bono, Armin Picenoni

Research Partner: University Hospital Zurich, Dr. Emanuela Felley-Bosco

Main Industrial Partner: FGen GmbH, Dr. Andreas Meyer, René Pellaux, Corinna Hund

Start: 01.01.2017, Duration: 24 months

Project goalThe screening of large cell libraries is an important process in

pharmaceutical discovery and R&D, e.g. to define drug targets or

develop effective medicines. The goal of this project is the

implementation of a screening platform based on 3D cultivation of

primary human mesothelioma cells encapsulated in alginate

hydrogels (Fig. 1) . To this end new hydrogel compositions will be

designed, tested and finally utilized in the Nanoliter Reactor

(NLR) cultivation system (Fig. 2) that enables high throughput

analysis of 3D cell cultures.

Key Findings• Successful manual cell encapsulation of mesothelioma cell line

ZL55SPT into alginate beads (Fig. 1A)

• Life/dead staining of encapsulated ZL55SPT cells (Fig. 1B)

Scientific Innovation• 3D cell culture of mammalian cells in Ca-alginate hydrogels

offering different microenvironments:

o Blending of alginate with natural environment mimicking

compounds

o Modification of the hydrogel stiffness to adapt to native

tissue

• Implementation of the modified Ca-alginate hydrogel in the

NLR-technology to produce reliable monodisperse capsules

at high reproducibility.

• Development of a novel method to generate patient-derived

primary mesothelioma cell lines in 3D (e.g. cell

subpopulations, like cancer stem cells) from a biopsy in a

high throughput manner using the NLR-technology.

• Retain the encapsulated mesothelioma cells in their native

state to use them for drug development/treatment to gain

predictive drug responses.

• Comparison of in vitro to in vivo and come up with

strategies to develop personalized drug treatment regimes.

Fig. 1: Mesothelioma cell line ZL55SPT encapsulated in alginate beads. A) Cell

growth and spheroid formation after 9 days of incubation. B) Life/Dead staining

(green/red) of cells after 21 days of incubation.

Fig. 2: NLR technology scheme

Cultivation106 NLRs/100 mL

Encapsulation0.7 - 3.5 kHz

Cell library

105 – 107 cells Ø: 20 – 600 µm

NLRs with cells

Phenotypic Assay

NLRs with clusters

>200 cells/cluster

Fluorescent hits

3 fluorescent signals

Characterization

Secondary assays

Analysis/Selection

10 – 250 Hz

Isolation from NLR

A B

brought to you by COREView metadata, citation and similar papers at core.ac.uk

provided by ZHAW digitalcollection