title: Probing the biology of cell boundary conditions through confinement of Xenopus cell-free cytoplasmic extracts.
abstract: Cell-free cytoplasmic extracts prepared from Xenopus eggs and embryos have for decades provided a biochemical system with which to interrogate complex cell biological processes in vitro. Recently, the application of microfabrication and microfluidic strategies in biology has narrowed the gap between in vitro and in vivo studies by enabling formation of cell-size compartments containing functional cytoplasm. These approaches provide numerous advantages over traditional biochemical experiments performed in a test tube. Most notably, the cell-free cytoplasm is confined using a two- or three-dimensional boundary, which mimics the natural configuration of a cell. This strategy enables characterization of the spatial organization of a cell, and the role that boundaries play in regulating intracellular assembly and function. In this review, we describe the marriage of Xenopus cell-free cytoplasm and confinement technologies to generate synthetic cell-like systems, the recent biological insights they have enabled, and the promise they hold for future scientific discovery.
authors.affiliation: Department of Bioengineering, University of Pennsylvania, 421 Curie Blvd, 1151 BRB II/III, Philadelphia, Pennsylvania, 19104.
authors.forename: Jessica G
authors.initials: JG
authors.lastname: Bermudez
authors.affiliation: Department of Cell and Developmental Biology, University of Pennsylvania, 421 Curie Blvd, 1151 BRB II/III, Philadelphia, Pennsylvania, 19104.
authors.forename: Hui
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Cell and Developmental Biology, University of Pennsylvania, 421 Curie Blvd, 1151 BRB II/III, Philadelphia, Pennsylvania, 19104.
authors.forename: Lily C
authors.initials: LC
authors.lastname: Einstein
authors.affiliation: Department of Bioengineering, University of Pennsylvania, 421 Curie Blvd, 1151 BRB II/III, Philadelphia, Pennsylvania, 19104.|Department of Cell and Developmental Biology, University of Pennsylvania, 421 Curie Blvd, 1151 BRB II/III, Philadelphia, Pennsylvania, 19104.
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Good
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002457:Cell Extracts / Q000235:genetics*; D002474:Cell-Free System; D003593:Cytoplasm / Q000235:genetics* / Q000378:metabolism; D044085:Microfluidics; D010063:Ovum / Q000254:growth & development / Q000378:metabolism; D014982:Xenopus laevis / Q000235:genetics*
chemical_list: D000199:Actins; D002457:Cell Extracts
keywords: Xenopus egg extract; cellular reconstitution; compartmentalization; encapsulation; microfluidics; synthetic cell