title: Three-dimensional scaffolding to investigate neuronal derivatives of human embryonic stem cells.
abstract: Access to unlimited numbers of live human neurons derived from stem cells offers unique opportunities for in vitro modeling of neural development, disease-related cellular phenotypes, and drug testing and discovery. However, to develop informative cellular in vitro assays, it is important to consider the relevant in vivo environment of neural tissues. Biomimetic 3D scaffolds are tools to culture human neurons under defined mechanical and physico-chemical properties providing an interconnected porous structure that may potentially enable a higher or more complex organization than traditional two-dimensional monolayer conditions. It is known that even minor variations in the internal geometry and mechanical properties of 3D scaffolds can impact cell behavior including survival, growth, and cell fate choice. In this report, we describe the design and engineering of 3D synthetic polyethylene glycol (PEG)-based and biodegradable gelatin-based scaffolds generated by a free form fabrication technique with precise internal geometry and elastic stiffnesses. We show that human neurons, derived from human embryonic stem (hESC) cells, are able to adhere to these scaffolds and form organoid structures that extend in three dimensions as demonstrated by confocal and electron microscopy. Future refinements of scaffold structure, size and surface chemistries may facilitate long term experiments and designing clinically applicable bioassays.
authors: Department of NanoEngineering, University of California, San Diego, 9500 Gilman Drive, Atkinson Hall, MC-0448, La Jolla, CA 92093, USA.
authors: Pranav
authors: P
authors: Soman
authors: Program in Stem Cell & Regenerative Biology, Sanford-Burnham Medical Research Institute, La Jolla, CA 92037, USA.
authors: Brian T D
authors: BTD
authors: Tobe
authors: Department of NanoEngineering, University of California, San Diego, 9500 Gilman Drive, Atkinson Hall, MC-0448, La Jolla, CA 92093, USA.
authors: Jin Woo
authors: JW
authors: Lee
authors: Program in Stem Cell & Regenerative Biology, Sanford-Burnham Medical Research Institute, La Jolla, CA 92037, USA.
authors: Alicia M
authors: AM
authors: Winquist
authors: Program in Stem Cell & Regenerative Biology, Sanford-Burnham Medical Research Institute, La Jolla, CA 92037, USA.
authors: Ilyas
authors: I
authors: Singec
authors: Department of NanoEngineering, University of California, San Diego, 9500 Gilman Drive, Atkinson Hall, MC-0448, La Jolla, CA 92093, USA.
authors: Kenneth S
authors: KS
authors: Vecchio
authors: Program in Stem Cell & Regenerative Biology, Sanford-Burnham Medical Research Institute, La Jolla, CA 92037, USA.
authors: Evan Y
authors: EY
authors: Snyder
authors: Department of NanoEngineering, University of California, San Diego, 9500 Gilman Drive, Atkinson Hall, MC-0448, La Jolla, CA 92093, USA.
authors: Shaochen
authors: S
authors: Chen
mesh_terms: D040761:Biomimetic Materials / Q000737:chemistry; D002448:Cell Adhesion; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000648:ultrastructure*; D004867:Equipment Design; D005780:Gelatin / Q000737:chemistry; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D009474:Neurons / Q000166:cytology*; D011092:Polyethylene Glycols / Q000737:chemistry; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D011092:Polyethylene Glycols; D005780:Gelatin