Degree Type

Thesis

Date of Award

2017

Degree Name

Master of Science

Department

Biomedical Sciences

Major

Biomedical Sciences

First Advisor

Donald S. Sakaguchi

Abstract

The ultimate purpose of this project was to create a modified stem cell line which could enhance nerve regeneration following peripheral nerve trauma. Specifically, this work was focused upon answering two questions. First, could we chemically transdifferentiate genetically modified mesenchymal stem cells to resemble a Schwann cell-like state? A protocol for the chemical transdifferentiation of MSCs was validated and well tested in the Sakaguchi lab, but no one had ever attempted to chemically transdifferentiate BDNF hyper-secreting MSCs. Second, if we succeeded in creating BDNF hyper-secreting transdifferentiated MSCs (BDNF tMSCs), would levels of BDNF secretion be affected, and, more importantly, would the secreted BDNF still be biologically active? We hypothesized that BDNF tMSCs would still resemble a Schwann cell like phenotype and be able to produce the same or lower amounts of biologically active BDNF when compared to their undifferentiated and GFP control counterparts. Generated data relied largely on the use of immunocytochemistry to quantify the percentage of cells expressing Schwann cell markers. BDNF secretion was quantified by ELISA and bioactivity was tested using the PC12-trkB assay. This study was an important first step in characterizing these BDNF tMSCs by in vitro assays and was essentially a proof of concept study to show that genetically modified MSCs could still be chemically transdifferentiated. As a next step, we hope to seed these BDNF tMSCs within a polymeric conduit transplant used in a rat sciatic nerve transection model to test the ability of these cells to aid in nerve regeneration in vivo.

DOI

https://doi.org/10.31274/etd-180810-4889

Copyright Owner

Metzere Bierlein De La Rosa

Language

en

File Format

application/pdf

File Size

90 pages

Included in

Cell Biology Commons

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