Gene-supplemented collagen scaffolds for non-viral gene delivery for brain tissue engineering

Recent advances in tissue engineering, combining an extracellular matrix (ECM)-like vehicle with therapeutic molecules, cells and/or genes has yielded promising results for brain injury repair. The purpose of this thesis was to develop a collagen scaffold for the non-viral delivery of the gene encoding for Glial Cell-Derived Neurotrophic Factor (GDNF); hence to provide a local, long-term release and overexpression of GDNF via transfection of cells seeded into the scaffold or endogenous cells. The first part of the thesis aimed to investigate the in vitro transfection of marrow stromal stem cells (also referred to as mesenchymal stem cells, MSCs) in monolayer with plasmid GDNF ([mu]gDNF). Several parameters were evaluated: the choice of a transfer reagent (GenePorter2 versus Lipofectamine 2000), the doses of plasmid incorporated in the liposomes (ranging from 0.2[mu]g to 2[mu]g), the post-transfection medium (Medium 1: DMEM low glucose, 20% FBS and 1% antibiotic versus Medium 2: DMEM low glucose, 20% FBS, 1% antibiotic and 10ng/ml FGF-2) and the culture environment during transfection (static versus dynamic). The objective of the second part was to determine the conditions, including the design of the scaffold and the method of seeding, under which MSCs could attach and grow on the scaffold.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Materials Science and Engineering.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2007

Author dc:creator

• Bolliet, Catherine

Advisor dc:contributor.advisor

• Myron Spector.

• Materials Science and Engineering.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/38586

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/38586