Fabrication of 3‐D Reconstituted Organoid Arrays by DNA‐Programmed Assembly of Cells (DPAC). Issue 3 (13th September 2016)

• Record Type: Journal Article
• Authors: Todhunter, Michael E.; Weber, Robert J.; Farlow, Justin; Jee, Noel Y.; Cerchiari, Alec E.; Gartner, Zev J.
• Editors: Mahal, Lara; Romesberg, Floyd; Shah, Kavita; Shamu, Caroline; Strano, Michael S.; Thomas, Craig
• Is Part Of: Current protocols in chemical biology. Volume 8:Issue 3(2016)
• Page Start: 147
• Page End: 178
• Publication Date: 2016-09-13
• Subjects: tissue array -- DNA -- synthetic biology -- cell‐cell interactions -- tissue engineering -- patterning -- organotypic -- organoid -- 3‐D culture; Biochemistry -- Laboratory manuals; Chemical Phenomena; Biochemistry; Periodicals; Laboratory manuals; 572.078
• DOI: 10.1002/cpch.8
• Languages: English
• ISSNs: 2160-4762

Abstract

Abstract: Tissues are the organizational units of function in metazoan organisms. Tissues comprise an assortment of cellular building blocks, soluble factors, and extracellular matrix (ECM) composed into specific three‐dimensional (3‐D) structures. The capacity to reconstitute tissues in vitro with the structural complexity observed in vivo is key to understanding processes such as morphogenesis, homeostasis, and disease. In this article, we describe DNA‐programmed assembly of cells (DPAC), a method to fabricate viable, functional arrays of organoid‐like tissues within 3‐D ECM gels. In DPAC, dissociated cells are chemically functionalized with degradable oligonucleotide "Velcro, " allowing rapid, specific, and reversible cell adhesion to a two‐dimensional (2‐D) template patterned with complementary DNA. An iterative assembly process builds up organoids, layer‐by‐layer, from this initial 2‐D template and into the third dimension. Cleavage of the DNA releases the completed array of tissues that are captured and fully embedded in ECM gels for culture and observation. DPAC controls the size, shape, composition, and spatial heterogeneity of organoids and permits positioning of constituent cells with single‐cell resolution even within cultures several centimeters long. © 2016 by John Wiley & Sons, Inc.