title: Transwells with microstamped membranes produce micropatterned two-dimensional and three-dimensional co-cultures.
abstract: This article describes a simple and rapid cell patterning method to form co-culture microarrays in commercially available Transwells. A thin poly(dimethylsiloxane) (PDMS) layer is printed on the underside of a Transwell using a PDMS stamp. Arbitrary cellular patterns are generated according to the geometric features of the thin PDMS layer through hydrodynamic forces that guide cells onto the membrane only over the PDMS-uncoated regions. Micropatterns of surface-adhered cells (we refer to this as two-dimensional) or non-surface-adhered clusters of cells (we refer to this as three-dimensional) can be generated depending on the surface treatment of the filter membrane. Additionally, co-cultures can be established by introducing different types of cells on the membrane or in the bottom chamber of the Transwell. We show that this co-culture method can evaluate mouse embryonic stem (mES) cell differentiation based on heterogeneous cell-cell interactions. Co-culture of mES cells and HepG2 cells decreased SOX17 expression of mES cells, and direct cell-cell contact further decreased SOX17 expression, indicating that co-culture with HepG2 cells inhibits endoderm differentiation through soluble factors and cell-cell contact. This method is simple and user-friendly and should be broadly useful to study cell shapes and cell-cell interactions.
authors.affiliation: 1 Department of Biomedical Engineering, University of Michigan , Ann Arbor, Michigan.
authors.forename: Yu-Suke
authors.initials: YS
authors.lastname: Torisawa
authors.forename: Bobak
authors.initials: B
authors.lastname: Mosadegh
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Cavnar
authors.forename: Mitchell
authors.initials: M
authors.lastname: Ho
authors.forename: Shuichi
authors.initials: S
authors.lastname: Takayama
mesh_terms: D000818:Animals; D019556:COS Cells; D002450:Cell Communication; D002454:Cell Differentiation; D002460:Cell Line; D048430:Cell Shape; D018920:Coculture Techniques / Q000295:instrumentation* / Q000379:methods; D003470:Culture Media; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D053595:Embryonic Stem Cells / Q000166:cytology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D024002:HMGB Proteins / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D057446:Hydrodynamics; D008422:Materials Testing; D051379:Mice; D055760:SOXF Transcription Factors / Q000378:metabolism; D013499:Surface Properties; D046888:Tissue Array Analysis; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods
chemical_list: D003470:Culture Media; D004129:Dimethylpolysiloxanes; D024002:HMGB Proteins; D055760:SOXF Transcription Factors; C511606:Sox17 protein, mouse; D049452:Green Fluorescent Proteins; C013830:baysilon