title: Layered patterning of hepatocytes in co-culture systems using microfabricated stencils.
abstract: Microfabrication and micropatterning techniques in tissue engineering offer great potential for creating and controlling microenvironments in which cell behavior can be observed. Here we present a novel approach to generate layered patterning of hepatocytes on micropatterned fibroblast feeder layers using microfabricated polydimethylsiloxane (PDMS) stencils. We fabricated PDMS stencils to pattern circular holes with diameters of 500 microm. Hepatocytes were co-cultured with 3T3-J2 fibroblasts in two types of patterns to evaluate and characterize the cellular interactions in the co-culture systems. Results of this study demonstrated uniform intracellular albumin staining and E-cadherin expression, increased liver-specific functions, and active glycogen synthesis in the hepatocytes when the heterotypic interface between hepatocytes and fibroblasts was increased by the layered patterning technique. This patterning technique can be a useful experimental tool for applications in basic science, drug screening, and tissue engineering, as well as in the design of bioartificial liver devices.
authors.affiliation: Center for Engineering in Medicine and Department of Surgery, Massachusetts General Hospital, Harvard Medical School, 51 Blossom St., Boston, MA 02114, USA. cho@njit.edu
authors.forename: Cheul H
authors.initials: CH
authors.lastname: Cho
authors.forename: Jaesung
authors.initials: J
authors.lastname: Park
authors.forename: Arno W
authors.initials: AW
authors.lastname: Tilles
authors.forename: François
authors.initials: F
authors.lastname: Berthiaume
authors.forename: Mehmet
authors.initials: M
authors.lastname: Toner
authors.forename: Martin L
authors.initials: ML
authors.lastname: Yarmush
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000295:instrumentation* / Q000379:methods; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004867:Equipment Design; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D055616:Microtechnology; D051381:Rats; D011917:Rats, Inbred Lew; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods
chemical_list: D004129:Dimethylpolysiloxanes; C013830:baysilon