DOI

10.17077/etd.5vjw-r15l

Document Type

Thesis

Date of Degree

Spring 2019

Degree Name

MS (Master of Science)

Degree In

Genetics

First Advisor

Kuehn, Markus

First Committee Member

Anderson, Michael

Second Committee Member

Grueter, Chad

Abstract

In the Kuehn lab, it has been shown that inducible pluripotent stems cells that have been induced to be trabecular meshwork cell-like (iPSC-TM) have a unique ability to regenerate dysfunctional trabecular meshwork (TM) cells by sharing specific unknown factors. In this thesis will discuss the novel means by which I isolate primary human Trabecular Meshwork (pTMs) and efficiently prepare cell cultures for experimentation, such as a sequencing experiment in which I studied expression changes that arose when the TM cell culture’s cell cycle control is manipulated. Previous research has shown that pTM grow atypical when 100% confluent compared to other epithelial cells creating an interesting time frame by which to observe their unique cell cycle control. Using newly isolated TM cell cultures I investigated expression of mRNA and miRNA to understand their roles in cell cycle control of these atypical cultures. With regards to the isolation of TM cell cultures were able to show that the “Crawling Out” methodology is an effective way to establish a pure TM cell line with both a low contamination rate and less passages/time. With these cultures we were able to establish 50 mRNAs and 19 miRNAs that were differential expressed in the TM cell cultures that were atypically grown. When reviewing the literature many of these expression changes were linked to carcinogenics, and the progression/prognosis of various cancer types.

Keywords

Cell Culture, Gene Expression, miRNA, Primary Open Angle Glaucoma, Trabecular Meshwork

Pages

vi, 27 pages

Bibliography

Includes bibliographical references (pages 26-27).

Copyright

Copyright © 2019 Kyle Joseph Gonsalves

Additional Files

supplemental fig 1.pdf (1330 kB)
Supplemental fig 2.pdf (31 kB)

Included in

Genetics Commons

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