Two and three-dimensional gene transfer from enzymatically degradable hydrogel scaffolds.
The ability to genetically modify mesenchymal stem cells (MSCs) seeded inside synthetic hydrogel scaffolds would offer an alternative approach to guide MSC differentiation. In this report, we explored gene transfer to MSCs seeded on top or inside matrix metalloproteinase (MMP) degradable hydrogels that were loaded with DNA/poly(ethylene imine) (PEI) polyplexes. DNA/PEI polyplexes were encapsulated inside poly(ethylene glycol) (PEG) hydrogels crosslinked with MMP degradable peptides via Michael Addition chemistry. Gene transfer was visualized and quantified through using a vector encoding for green fluorescent protein and luciferase. We found that gene transfer to MSCs was possible for cells seeded both in two and three dimensions. The amount of luciferase expression was similar for cells seeded in two and three dimensions even though the number of cells in three dimensions is significantly higher, indicating that gene transfer to cells seeded in two dimensions is more efficient than for cells seeded in three dimensions. The use of hydrogel scaffolds that allow cellular infiltration to deliver DNA may result in long-lasting signals in vivo, which are essential for the regeneration of functional tissues.
Chemical and Biomolecular Engineering Department, University of California, Los Angeles, California, USA.
Yuguo
Y
Lei
Quinn K T
QK
Ng
Tatiana
T
Segura
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D004247:DNA / Q000737:chemistry / Q000235:genetics; D018014:Gene Transfer Techniques / Q000295:instrumentation*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D006868:Hydrolysis; D007097:Imines / Q000737:chemistry; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000737:chemistry*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D051379:Mice; D011095:Polyethylenes / Q000737:chemistry; D023822:Tissue Engineering / Q000295:instrumentation*; D054457:Tissue Scaffolds / Q000737:chemistry*
D007097:Imines; D011095:Polyethylenes; C505405:poly(ethylene imine); D049452:Green Fluorescent Proteins; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D004247:DNA; D008156:Luciferases; D020780:Matrix Metalloproteinase 9