title: Electrospun gelatin biopapers as substrate for in vitro bilayer models of blood-brain barrier tissue.
abstract: Gaining a greater understanding of the blood-brain barrier (BBB) is critical for improvement in drug delivery, understanding pathologies that compromise the BBB, and developing therapies to protect the BBB. In vitro human tissue models are valuable tools for studying these issues. The standard in vitro BBB models use commercially available cell culture inserts to generate bilayer co-cultures of astrocytes and endothelial cells (EC). Electrospinning can be used to produce customized cell culture substrates with optimized material composition and mechanical properties with advantages over off-the-shelf materials. Electrospun gelatin is an ideal cell culture substrate because it is a natural polymer that can aid cell attachment and be modified and degraded by cells. Here, we have developed a method to produce cell culture inserts with electrospun gelatin "biopaper" membranes. The electrospun fiber diameter and cross-linking method were optimized for the growth of primary human endothelial cell and primary human astrocyte bilayer co-cultures to model human BBB tissue. BBB co-cultures on biopaper were characterized via cell morphology, trans-endothelial electrical resistance (TEER), and permeability to FITC-labeled dextran and compared to BBB co-cultures on standard cell culture inserts. Over longer culture periods (up to 21 days), cultures on the optimized electrospun gelatin biopapers were found to have improved TEER, decreased permeability, and permitted a smaller separation between co-cultured cells when compared to standard PET inserts.
authors.affiliation: American Society for Engineering Education Postdoctoral Fellow at the U.S. Naval Research Laboratory, Washington, DC.
authors.forename: Lauren L
authors.initials: LL
authors.lastname: Bischel
authors.affiliation: American Society for Engineering Education Postdoctoral Fellow at the U.S. Naval Research Laboratory, Washington, DC.
authors.forename: Peter N
authors.initials: PN
authors.lastname: Coneski
authors.affiliation: U.S. Naval Research Laboratory, Chemistry Division, Washington, DC.
authors.forename: Jeffrey G
authors.initials: JG
authors.lastname: Lundin
authors.affiliation: Department of Physics, Southern Oregon University, Ashland, Oregon.
authors.forename: Peter K
authors.initials: PK
authors.lastname: Wu
authors.affiliation: Contractor at the U.S. Naval Research Laboratory, Leidos, Washington, DC.
authors.forename: Carl B
authors.initials: CB
authors.lastname: Giller
authors.affiliation: U.S. Naval Research Laboratory, Chemistry Division, Washington, DC.
authors.forename: James
authors.initials: J
authors.lastname: Wynne
authors.affiliation: U.S. Naval Research Laboratory, Chemistry Division, Washington, DC.
authors.forename: Brad R
authors.initials: BR
authors.lastname: Ringeisen
authors.affiliation: U.S. Naval Research Laboratory, Chemistry Division, Washington, DC.
authors.forename: Russell K
authors.initials: RK
authors.lastname: Pirlo
mesh_terms: D001253:Astrocytes / Q000166:cytology*; D001672:Biocompatible Materials / Q000737:chemistry*; D001812:Blood-Brain Barrier / Q000166:cytology*; D001921:Brain / Q000166:cytology; D002460:Cell Line; D018920:Coculture Techniques / Q000379:methods; D003432:Cross-Linking Reagents / Q000737:chemistry; D004560:Electricity; D042783:Endothelial Cells / Q000166:cytology*; D005780:Gelatin / Q000737:chemistry*; D006801:Humans; D008567:Membranes, Artificial; D010209:Paper
chemical_list: D001672:Biocompatible Materials; D003432:Cross-Linking Reagents; D008567:Membranes, Artificial; D005780:Gelatin
keywords: biomaterial; blood-brain barrier; electrospinning; gelatin; tissue engineering