Fabrication of 3-D Reconstituted Organoid Arrays by DNA-Programmed Assembly of Cells (DPAC).
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.
Department of Pharmaceutical Chemistry, University of California, San Francisco, California.|Tetrad Graduate Program, University of California, San Francisco, California.
Michael E
ME
Todhunter
Department of Pharmaceutical Chemistry, University of California, San Francisco, California.|Chemistry & Chemical Biology Graduate Program, University of California, San Francisco, California.
Robert J
RJ
Weber
Department of Pharmaceutical Chemistry, University of California, San Francisco, California.|Tetrad Graduate Program, University of California, San Francisco, California.
Justin
J
Farlow
Department of Pharmaceutical Chemistry, University of California, San Francisco, California.|Chemistry & Chemical Biology Graduate Program, University of California, San Francisco, California.
Noel Y
NY
Jee
Department of Pharmaceutical Chemistry, University of California, San Francisco, California.|Graduate Program in Bioengineering, University of California, Berkeley, and University of California, San Francisco, California.
Alec E
AE
Cerchiari
Department of Pharmaceutical Chemistry, University of California, San Francisco, California.|Center for Systems and Synthetic Biology, University of California, San Francisco, California.
Zev J
ZJ
Gartner
D000067470:Cellular Reprogramming Techniques / Q000379:methods*; D004247:DNA / Q000737:chemistry*; D006801:Humans; D009940:Organoids / Q000737:chemistry*; D023822:Tissue Engineering / Q000379:methods*
D004247:DNA
3-D culture; DNA; cell-cell interactions; organoid; organotypic; patterning; synthetic biology; tissue array; tissue engineering