title: Seamless Combination of Fluorescence-Activated Cell Sorting and Hanging-Drop Networks for Individual Handling and Culturing of Stem Cells and Microtissue Spheroids.
abstract: Open microfluidic cell culturing devices offer new possibilities to simplify loading, culturing, and harvesting of individual cells or microtissues due to the fact that liquids and cells/microtissues are directly accessible. We present a complete workflow for microfluidic handling and culturing of individual cells and microtissue spheroids, which is based on the hanging-drop network concept: The open microfluidic devices are seamlessly combined with fluorescence-activated cell sorting (FACS), so that individual cells, including stem cells, can be directly sorted into specified culturing compartments in a fully automated way and at high accuracy. Moreover, already assembled microtissue spheroids can be loaded into the microfluidic structures by using a conventional pipet. Cell and microtissue culturing is then performed in hanging drops under controlled perfusion. On-chip drop size control measures were applied to stabilize the system. Cells and microtissue spheroids can be retrieved from the chip by using a parallelized transfer method. The presented methodology holds great promise for combinatorial screening of stem-cell and multicellular-spheroid cultures.
authors.affiliation: ETH Zurich , Department of Biosystems Science and Engineering, Bio Engineering Laboratory, Mattenstrasse 26, CH-4058 Basel, Switzerland.
authors.forename: Axel
authors.initials: A
authors.lastname: Birchler
authors.affiliation: ETH Zurich , Department of Biosystems Science and Engineering, Bio Engineering Laboratory, Mattenstrasse 26, CH-4058 Basel, Switzerland.
authors.forename: Mischa
authors.initials: M
authors.lastname: Berger
authors.affiliation: ETH Zurich , Department of Biosystems Science and Engineering, Single Cell Facility, Mattenstrasse 26, CH-4058 Basel, Switzerland.
authors.forename: Verena
authors.initials: V
authors.lastname: Jäggin
authors.affiliation: ETH Zurich , Department of Biosystems Science and Engineering, Single Cell Facility, Mattenstrasse 26, CH-4058 Basel, Switzerland.
authors.forename: Telma
authors.initials: T
authors.lastname: Lopes
authors.affiliation: ETH Zurich , Department of Biosystems Science and Engineering, Cell Systems Dynamics Group, Mattenstrasse 26, CH-4058 Basel, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Etzrodt
authors.affiliation: ETH Zurich , Department of Biosystems Science and Engineering, Bio Engineering Laboratory, Mattenstrasse 26, CH-4058 Basel, Switzerland.
authors.forename: Patrick Mark
authors.initials: PM
authors.lastname: Misun
authors.affiliation: ETH Zurich , Department of Biosystems Science and Engineering, Bio Engineering Laboratory, Mattenstrasse 26, CH-4058 Basel, Switzerland.
authors.forename: Maria
authors.initials: M
authors.lastname: Pena-Francesch
authors.affiliation: ETH Zurich , Department of Biosystems Science and Engineering, Cell Systems Dynamics Group, Mattenstrasse 26, CH-4058 Basel, Switzerland.
authors.forename: Timm
authors.initials: T
authors.lastname: Schroeder
authors.affiliation: ETH Zurich , Department of Biosystems Science and Engineering, Bio Engineering Laboratory, Mattenstrasse 26, CH-4058 Basel, Switzerland.
authors.forename: Andreas
authors.initials: A
authors.lastname: Hierlemann
authors.affiliation: ETH Zurich , Department of Biosystems Science and Engineering, Bio Engineering Laboratory, Mattenstrasse 26, CH-4058 Basel, Switzerland.
authors.forename: Olivier
authors.initials: O
authors.lastname: Frey
mesh_terms: D018929:Cell Culture Techniques; D005434:Flow Cytometry; D045325:HCT116 Cells; D006801:Humans; D046210:Microfluidic Analytical Techniques; D010316:Particle Size; D018874:Spheroids, Cellular / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*