title: Genetically encoded sender-receiver system in 3D mammalian cell culture.
abstract: Engineering spatial patterning in mammalian cells, employing entirely genetically encoded components, requires solving several problems. These include how to code secreted activator or inhibitor molecules and how to send concentration-dependent signals to neighboring cells, to control gene expression. The Madin-Darby Canine Kidney (MDCK) cell line is a potential engineering scaffold as it forms hollow spheres (cysts) in 3D culture and tubulates in response to extracellular hepatocyte growth factor (HGF). We first aimed to graft a synthetic patterning system onto single developing MDCK cysts. We therefore developed a new localized transfection method to engineer distinct sender and receiver regions. A stable reporter line enabled reversible EGFP activation by HGF and modulation by a secreted repressor (a truncated HGF variant, NK4). By expanding the scale to wide fields of cysts, we generated morphogen diffusion gradients, controlling reporter gene expression. Together, these components provide a toolkit for engineering cell-cell communication networks in 3D cell culture.
authors: EMBL/CRG Systems Biology Research Unit, Centre for Genomic Regulation (CRG) , Dr. Aiguader 88, 08003 Barcelona, Spain.
authors: Andreia
authors: A
authors: Carvalho
authors: Diego Barcena
authors: DB
authors: Menendez
authors: Vivek Raj
authors: VR
authors: Senthivel
authors: Timo
authors: T
authors: Zimmermann
authors: Luis
authors: L
authors: Diambra
authors: Mark
authors: M
authors: Isalan
mesh_terms: D000818:Animals; D002450:Cell Communication / Q000235:genetics* / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods*; D060846:Cell Engineering / Q000379:methods*; D004285:Dogs; D017228:Hepatocyte Growth Factor / Q000235:genetics / Q000378:metabolism; D061985:Madin Darby Canine Kidney Cells; D015398:Signal Transduction / Q000235:genetics* / Q000502:physiology; D058615:Synthetic Biology / Q000379:methods*
chemical_list: D017228:Hepatocyte Growth Factor