NOVEL 3D CELL CULTURE TECHNIQUE UTILIZING A SILICON TITANIUM DIBORIDE MICROPATTERNED SUBSTRATE FOR DIFFERENTIATING MESENCHYMAL STEM CELLS INTO INSULIN PRODUCING CELLS

dc.contributor.advisorMerchant, Fatima Aziz
dc.contributor.committeeMemberAkay, Yasemin M.
dc.contributor.committeeMemberWu, Tianfu
dc.contributor.committeeMemberAl-Ubaidi, Muayyad R.
dc.contributor.committeeMemberSabek, Omaima
dc.creatorFriguglietti, Jefferson
dc.creator.orcid0000-0001-8175-635X
dc.date.accessioned2024-01-26T17:45:16Z
dc.date.createdDecember 2023
dc.date.issued2023-12
dc.date.updated2024-01-26T17:45:17Z
dc.description.abstract3D cell culture techniques are increasingly used in stem cell tissue engineering since they better mimic the in vivo environments compared to conventional 2D culture. Current 3D culture methods include less complex suspension methods such as hanging well and ultra-low attachment plate, to more advance methods incorporating scaffold designs to support cell-cell and cell-extracellular matrix interaction (ECM). In this study we investigate a novel microfabricated silicon-titanium diboride (Si-TiB2) substrate’s efficacy in increasing function of differentiated mesenchymal stem cells (MSCs) into insulin producing cells (IPCs). This substrate provides a 3D microenvironment (aggregates) and geometric (pattern shape), mechanical (stiffness gradients) and biochemical (selectively adsorbed proteins) cues that are critical for cell differentiation. Differentiated IPCs are essential, providing an alternative tissue bank for future transplantation into diabetic patients. Utilizing photolithography technique, e-beam deposited TiB2 layers were fabricated on Si, wherein differences in their surface properties (hardness, stiffness, wetness, and electrical charge), enable selective adsorption of specific proteins on the micropatterns. MSCs from adult bone marrow were cultured on the substrate, and following a seven-day culture period subjected to a differentiation protocol. Functional analysis included quantification of c-peptide following a glucose stimulated insulin response (GSIR) assay. Morphological analysis of the cytoskeleton through immunofluorescence staining (f-actin, green) revealed increased rounded morphology of cells within the multicellular aggregates confirming a 3D culture environment on the SiTiB2 substrate. Additionally, immunofluorescence staining for biomarkers (n - cadherin, red) showed increased expression within the rounded core indicating cell-cell interaction with the aggregates. Further BioAFM analysis indicated cells with higher elastic modulus in the center of aggregates compared to cells on the edges of the micropatterns (p < 0.05). GSIR assay showed MSCs differentiated on the Si-TiB2 substrate had a better response to high glucose stimulation when compared to those differentiated in 2D monolayers in tissue culture plates (p < 0.05). Finally, co – culture with HUVEC and hBM-MSCs revealed unique formation of aggregation with HUVEC located primarily in the center of the aggregate. The TiB2 substrate provides a unique culture platform to better understand differentiation of MSCs into insulin producing cells when compared to ULP and traditional 2D flask.
dc.description.departmentBiomedical Engineering, Department of
dc.format.digitalOriginborn digital
dc.format.mimetypeapplication/pdf
dc.identifier.citationPortions of this document appear in: Friguglietti, Jefferson, Susmi Das, Phi Le, Daniel Fraga, Marcos Quintela, Salvatore A. Gazze, Darius McPhail et al. "Novel silicon titanium diboride micropatterned substrates for cellular patterning." Biomaterials 244 (2020): 119927.
dc.identifier.urihttps://hdl.handle.net/10657/16180
dc.language.isoeng
dc.rightsThe author of this work is the copyright owner. UH Libraries and the Texas Digital Library have their permission to store and provide access to this work. UH Libraries has secured permission to reproduce any and all previously published materials contained in the work. Further transmission, reproduction, or presentation of this work is prohibited except with permission of the author(s).
dc.subjectTissue Engineering, 3D culture technique, Insulin, Patterned, TIB2 substrate, MSCs, Stem Cells, HUVEC, Glucose, Hardness, Stiffness, Photolithography, Differentiation, Silicon, Titanium
dc.titleNOVEL 3D CELL CULTURE TECHNIQUE UTILIZING A SILICON TITANIUM DIBORIDE MICROPATTERNED SUBSTRATE FOR DIFFERENTIATING MESENCHYMAL STEM CELLS INTO INSULIN PRODUCING CELLS
dc.type.dcmitext
dc.type.genreThesis
dcterms.accessRightsThe full text of this item is not available at this time because the student has placed this item under an embargo for a period of time. The Libraries are not authorized to provide a copy of this work during the embargo period.
local.embargo.lift2025-12-01
local.embargo.terms2025-12-01
thesis.degree.collegeCullen College of Engineering
thesis.degree.departmentBiomedical Engineering, Department of
thesis.degree.disciplineBiomedical Engineering
thesis.degree.grantorUniversity of Houston
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy

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