Biomedical Sciences ETDs

Publication Date

Summer 8-1-2023

Abstract

Development of a cell culture based bone model can contribute to biomedical research by producing a reproducible model system that mimics a bone remodeling disorder and reduces the use of animals in research studies. Synthetic biology was used to engineer osteoblast cell lines to alter a key protein in the RANKL pathway in efforts to control the bone remodeling process. Osteoblast cell lines, K7M2 and MC3T3-E1 Subclone 4 were engineered to express osteoprotegerin (OPG) and inhibit OPG expression, respectively. The upregulation of OPG expression was confirmed with Real Time PCR and Quantikine® Immunoassay. To identify the functionality of the engineered K7M2-OPG-Myc cell line, a resorption pit assay using calcium phosphate as synthetic biomimetic material was used. RAW cells were differentiated to osteoclasts using varying concentrations of RANKL in conditioned medium from engineered K7M2-OPG-Myc cell line in the calcium phosphate assay. Conditioned Medium from K7M2-OPG-Myc cells inhibited resorption activity of formed osteoclasts. This thesis work was the first step taken to enable the development of a cell culture based bone model capable of remodeling on its own.

Keywords

Osteoblasts, Osteoclasts, Osteocytes, Bone Remodeling Model

Document Type

Thesis

Language

English

Degree Name

Biomedical Sciences

Level of Degree

Masters

Department Name

Biomedical Sciences Graduate Program

First Committee Member (Chair)

Laura Gonzalez-Bosc

Second Committee Member

Christina Salas

Third Committee Member

Jerilyn A. Timlin

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