title: Promoting 3-D Aggregation of FACS Purified Thymic Epithelial Cells with EAK 16-II/EAKIIH6 Self-assembling Hydrogel.
abstract: Thymus involution, associated with aging or pathological insults, results in diminished output of mature T-cells. Restoring the function of a failing thymus is crucial to maintain effective T cell-mediated acquired immune response against invading pathogens. However, thymus regeneration and revitalization proved to be challenging, largely due to the difficulties of reproducing the unique 3D microenvironment of the thymic stroma that is critical for the survival and function of thymic epithelial cells (TECs). We developed a novel hydrogel system to promote the formation of TEC aggregates, based on the self-assembling property of the amphiphilic EAK16-II oligopeptides and its histidinylated analogue EAKIIH6. TECs were enriched from isolated thymic cells with density-gradient, sorted with fluorescence-activated cell sorting (FACS), and labeled with anti-epithelial cell adhesion molecule (EpCAM) antibodies that were anchored, together with anti-His IgGs, on the protein A/G adaptor complexes. Formation of cell aggregates was promoted by incubating TECs with EAKIIH6 and EAK16-II oligopeptides, and then by increasing the ionic concentration of the medium to initiate gelation. TEC aggregates embedded in EAK hydrogel can effectively promote the development of functional T cells in vivo when transplanted into the athymic nude mice.
authors.affiliation: Institute of Cellular Therapeutics, Allegheny Health Network.
authors.forename: Asako
authors.initials: A
authors.lastname: Tajima
authors.affiliation: Division of Pharmaceutical Sciences, Mylan School of Pharmacy, Duquesne University.
authors.forename: Wen
authors.initials: W
authors.lastname: Liu
authors.affiliation: Institute of Cellular Therapeutics, Allegheny Health Network.
authors.forename: Isha
authors.initials: I
authors.lastname: Pradhan
authors.affiliation: Institute of Cellular Therapeutics, Allegheny Health Network.
authors.forename: Suzanne
authors.initials: S
authors.lastname: Bertera
authors.affiliation: Institute of Cellular Therapeutics, Allegheny Health Network.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Lakomy
authors.affiliation: Institute of Cellular Therapeutics, Allegheny Health Network.
authors.forename: William A
authors.initials: WA
authors.lastname: Rudert
authors.affiliation: Institute of Cellular Therapeutics, Allegheny Health Network; Department of Biological Sciences, Carnegie Mellon University.
authors.forename: Massimo
authors.initials: M
authors.lastname: Trucco
authors.affiliation: Division of Pharmaceutical Sciences, Mylan School of Pharmacy, Duquesne University.
authors.forename: Wilson S
authors.initials: WS
authors.lastname: Meng
authors.affiliation: Institute of Cellular Therapeutics, Allegheny Health Network; Department of Biological Sciences, Carnegie Mellon University; yong.fan74@gmail.com.
authors.forename: Yong
authors.initials: Y
authors.lastname: Fan
mesh_terms: D000818:Animals; D004847:Epithelial Cells; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D051379:Mice; D008819:Mice, Nude; D009842:Oligopeptides; D013601:T-Lymphocytes; D013950:Thymus Gland
chemical_list: D009842:Oligopeptides; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate