Award Date

5-1-2022

Degree Type

Thesis

Degree Name

Master of Science (MS)

Department

Dental Medicine

First Committee Member

Katherine Howard

Second Committee Member

Karl Kingsley

Third Committee Member

Brian Chrzan

Fourth Committee Member

Courtney Coughenour

Number of Pages

70

Abstract

Introduction: Dental pulp stem cells (DPSC) are pluripotent stem cells capable of differentiating into several tissue types. Several studies have demonstrated DPSC can be extracted from deciduous and permanent teeth, including third molars (wisdom teeth) - however, much less is known regarding the factors that determine and regulate pluripotency and differentiation responsiveness among these harvested DPSC. Recent evidence has suggested small, non-coding microRNAs mediate (in part) differentiation and self-renewal among some stromal and hematopoetic stem cells, although much remains to be discovered regarding microRNA regulation of DPSC phenotypes. Based upon this information, the overall goal of this study was to determine if DPSC microRNA expression in exosomes correlates with DPSC phenotypes, such as differentiation status or growth. Methods: Six dental pulp stem cell (DPSC) isolates were retrieved from an existing repository and cultured. The original protocol for extraction and isolation of the DPSC was approved by the UNLV Biomedical IRB. Cells transferred into exosome-free media for 24 hours prior to exosome isolation and harvesting. Exosomes were analyzed using Particle Metrix Zeta view, Western blot analysis and qPCR screening. Results: All six DPSC cell lines were successfully established and cultured. Extracted exosome particle sizes ranged between 50 – 250 nm, corresponding with known exosome sizes. Bradford protein assays and Western Blots confirmed proteins (CD63) from isolated exosomes. RNA extraction and cDNA synthesis was performed in preparation for qPCR screening for microRNA expression with qPCR screening currently underway. Conclusions: These data confirm the successful isolation and characterization of DPSC exosomes and the corresponding RNA. qPCR screening revealed no correlation between miRNA expression and growth or differentiation status. Keywords: DPSC, miRNA, Exosomes

Keywords

DPSC; Exosomes; MiRNA

Disciplines

Dentistry

File Format

pdf

File Size

1602 KB

Degree Grantor

University of Nevada, Las Vegas

Language

English

Rights

IN COPYRIGHT. For more information about this rights statement, please visit http://rightsstatements.org/vocab/InC/1.0/

Download


Included in

Dentistry Commons

Share

COinS