Title | Transwells with microstamped membranes produce micropatterned two-dimensional and three-dimensional co-cultures. |
Publication Type | Journal Article |
Year of Publication | 2011 |
Authors | Torisawa Y-S, Mosadegh B, Cavnar SP, Ho M, Takayama S |
Journal | Tissue Eng Part C Methods |
Volume | 17 |
Issue | 1 |
Pagination | 61-7 |
Date Published | 2011 Jan |
ISSN | 1937-3392 |
Keywords | Animals, Cell Communication, Cell Differentiation, Cell Line, Cell Shape, Coculture Techniques, COS Cells, Culture Media, Dimethylpolysiloxanes, Embryonic Stem Cells, Green Fluorescent Proteins, Hep G2 Cells, HMGB Proteins, Humans, Hydrodynamics, Materials Testing, Mice, SOXF Transcription Factors, Surface Properties, Tissue Array Analysis, Tissue Engineering |
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. |
DOI | 10.1089/ten.TEC.2010.0305 |
Alternate Journal | Tissue Eng Part C Methods |
PubMed ID | 20673133 |
PubMed Central ID | PMC3011919 |
Grant List | R01CA136829-02 / CA / NCI NIH HHS / United States R01HL084370-05 / HL / NHLBI NIH HHS / United States |
Related Institute:
Dalio Institute of Cardiovascular Imaging (Dalio ICI)