2026-09-21T13:10:44Z https://www.ideals.illinois.edu/oai-pmh

oai:www.ideals.illinois.edu:2142/44809 2023-07-11 col_2142_5163 col_2142_5131 com_2142_5162 com_2142_234 com_2142_5130

2013-05 Cheng, Jianjun Wang, Fei Yen, Jonathan 2013-05-28T19:20:45Z 2013-05-28T19:20:45Z 2015-05-28T10:01:46Z 2013-05-28T19:20:45Z 2013-05 Gene delivery is an important tool to study and manipulate human pluripotent stem cells for regenerative medicine purposes. Yet current methods of transient gene delivery are still highly inefficient. Using materials and biologically based concepts, we aim to develop new methods and protocols to enhance the efficiency of gene delivery to be useful. For the materials aspect, diblock copolymers consisting of poly(ethylene glycol)-block-poly(γ-4-(((2-(piperidin-1-yl)ethyl)amino)methyl)benzyl-L-glutamate) (PEG-b-PVBLG-8) were synthesized and evaluated for their ability to mediate gene delivery in hard-to-transfect cells like IMR-90 human fetal lung fibroblasts and human embryonic stem cells (hESCs). The PEG-b-PVBLG-8 contained a membrane-disruptive, cationic, helical polypeptide block (PVBLG-8) for complexing with DNA and a hydrophilic PEG block to improve the biocompatibility of the gene delivery vehicle. PEG-b-PVBLG-8 diblock polymers with a high degree of polymerization have a greater transfection efficiency and lower toxicity in IMR-90 cells than the commercial reagent Lipofectamine 2000. The usefulness of PEG-b-PVBLG-8 was further demonstrated via the successful transfection of hESCs without a measured loss in cell pluripotency markers. From the biological aspect, a small molecule that selectively inhibits the Rho-associated kinase inhibitor (Y-27632) was discovered that transiently alters the hESC morphology to induce spreading and reduced membrane tension. These morphological changes allowed the increase of plasmid transfection, siRNA transfection and nanoparticle uptake to increase substantially. Treating the cells with Y-27632 and passaging them as single cells, we were able to obtain a transfection efficiency of ~90% in hESCs. Cells were also able to recover after treatment back to normal pluripotent stem cell morphology and express important pluripotency markers. Item withdrawn by Mark Zulauf (zulauf@illinois.edu) on 2013-04-24T18:46:12Z Item was in collections: University of Illinois Theses & Dissertations (ID: 1) No. of bitstreams: 4 Yen_Jonathan.pdf: 1847415 bytes, checksum: cab7d78929f603ab74376806e87677c3 (MD5) Yen_Jonathan.pdf: 1847178 bytes, checksum: 129db620a4ec50de3549d7cf79bbb3b9 (MD5) Yen_Jonathan.docx: 13386808 bytes, checksum: 4cc658bd4786caec17c872f7d83d4a3e (MD5) Yen_Jonathan.pdf: 1847415 bytes, checksum: cab7d78929f603ab74376806e87677c3 (MD5) Made available in DSpace on 2013-05-28T19:20:45Z (GMT). No. of bitstreams: 2 Jonathan_Yen.pdf: 1823920 bytes, checksum: a974559bbf4b0000a31823172900bff5 (MD5) license.txt: 4059 bytes, checksum: 019c33d62ac7935dec3365a571167b95 (MD5) Item marked as restricted to the 'Administrator' Group (id=1) by Seth Robbins (srobbins@illinois.edu) on 2013-05-28T19:22:03Z Item is restricted until 2015-05-28T19:21:22Z Restriction data tranferred 2014-07-01T11:17:20-05:00 Original Data Group with Access Administrator Release Date: 2015-05-28 14:21:22 UTC Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system Limited Restriction Lifted for Item 44784 on 2015-05-28T10:01:46Z. http://hdl.handle.net/2142/44809 en Copyright 2013 Jonathan Yen Stem Cells gene delivery human embryonic stem cells pluripotent stem cells peptides helical peptides fibroblasts small molecule Materials and biological approach to gene delivery in human embryonic stem cells text Bioengineering 1343 Bioengineering 0408 University of Illinois at Urbana-Champaign Thesis M.S. PHD: Bioengineering-UIUC 10KS0408PHD