Development of 3D hydrogel culture systems with on-demand cell separation

• Type of Material: Article
• Authors: Sharon K. Hamilton, Georgia Institute of Technology; Nathaniel C. Bloodworth, Georgia Institute of Technology; Christopher S. Massad, Georgia Institute of Technology; Taymour M. Hammoudi, Georgia Institute of Technology; Shalu Suri, Georgia Institute of Technology; Peter J. Yang, Georgia Institute of Technology; Hang Lu, Georgia Institute of Technology; Johnna Temenoff, Emory University
• Language: English
• Date: 2013-04-01
• Publisher: Wiley: 12 months
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
• Final Published Version (URL): http://dx.doi.org/10.1002/biot.201200200
• Title of Journal or Parent Work: Biotechnology Journal
• ISSN: 1860-6768
• Volume: 8
• Issue: 4
• Start Page: 485
• End Page: +
• Grant/Funding Information: The authors acknowledge funding from NIH/NIBIB (R21EB009153), NIH MSTP T32 GM008169, NIH ARRA (RC1CA144825), and the Petit Scholars Program; The hMSCs used in this work were provided by the Texas A&M Health Science Center College of Medicine Institute for Regenerative Medicine at Scott & White through a grant from NCRR of the NIH, Grant # P40RR017447.
• Keywords: Micropatterning; Biochemical Research Methods; ALGINATE HYDROGELS; Science & Technology; MESENCHYMAL STEM-CELLS; Hydrogel; OSTEOGENIC DIFFERENTIATION; Poly(ethylene glycol); Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; STROMAL CELLS; IN-VITRO; TISSUE; Co-culture; POLY(VINYL ALCOHOL); SOFT LITHOGRAPHY; POLY(ETHYLENE GLYCOL) HYDROGELS; Life Sciences & Biomedicine; Mesenchymal stem cell; MICROFLUIDIC DEVICES
• Research Categories: Engineering, Biomedical

Abstract

Recently there has been an increased interest in the effects of paracrine signaling between groups of cells, particularly in the context of better understanding how stem cells contribute to tissue repair. Most current 3D co-culture methods lack the ability to effectively separate two cell populations after the culture period, which is important for simultaneously analyzing the reciprocal effects of each cell type on the other. Here, we detail the development of a 3D hydrogel co-culture system that allows us to culture different cell types for up to 7 days and subsequently separate and isolate the different cell populations using enzyme-sensitive glues. Separable 3D co-culture laminates were prepared by laminating PEG-based hydrogels with enzyme-degradable hydrogel adhesives. Encapsulated cell populations exhibited good segregation with well-defined interfaces. Furthermore, constructs can be separated on-demand upon addition of the appropriate enzyme, while cell viability remains high throughout the culture period, even after laminate separation. This platform offers great potential for a variety of basic cell signaling studies as the incorporation of an enzyme-sensitive adhesive interface allows the on-demand separation of individual cell populations for immediate analysis or further culture to examine persistence of co-culture effects and paracrine signaling on cell populations.