Molecular, Cellular, and Tissue Engineering Commons^™

The Impact Of Dipeptides On Product Quality In Chinese Hamster Ovary Cells, Louise C. Herman

All Theses

Biopharmaceuticals are medicines produced in cells which are used to treat diseases such as cancer. Monoclonal antibodies (mAbs) represent the majority of approved biotherapeutics. MAbs are mostly produce using Chinese hamster ovary (CHO) cells. CHO cells are favored for resiliency, scalability, and established safety, and are considered the “workhorse” organism for biopharmaceuticals.  Amino acids are a key component of cell culture media, however several amino acids have solubility limits, such as cysteine, cystine, and tyrosine.  At neutral pH, these amino acids are poorly soluble.  However, dipeptides, such as N,N'-di-L-Alanyl-L-Cystine (ACCA) have been reported to increase cysteine and cystine availability. Further, …

Fluorescence-Based Quantification Of Hepatic Ldl Uptake And Donor Hepatocyte Engraftment Using Atto 700–Labeled Ldl, Victor Agustin Cortes Flores

All Theses

Familial hypercholesterolemia (FH) is an inherited disorder characterized by elevated low-density lipoprotein cholesterol (LDL-C) that increases the risk of premature atherosclerotic cardiovascular disease. In severe FH, including low-density lipoprotein receptor (LDLR), hepatic LDL clearance is profoundly impaired and many lipid lowering treatment strategies have limited effectiveness. Current advances in ex vivo CRISPR–Cas9 hepatocyte gene editing in vivo and ex vivo approaches for treatment of FH are under development.

However, a central barrier to translation is the ability to quantitatively measure the percentage of healthy gen edited hepatocytes in the liver while simultaneously assessing functional hepatic LDL handling. This thesis establishes …

Diabetes-Induced Mitral Valve Degeneration: Tissue-Engineered Solutions And Antioxidant Therapy, Samantha Hahn

All Theses

Mitral valve (MV) fibrosis is a progressive pathological condition characterized by leaflet thickening and stiffening, ultimately compromising valve function. This process is accelerated in diabetes, where chronic hyperglycemia elevates reactive oxygen species (ROS) and alters the mechanics of the extracellular matrix (ECM), collectively promoting the activation of valvular interstitial cells (VICs). Under healthy conditions, VICs maintain a quiescent, fibroblast-like phenotype responsible for sustaining ECM homeostasis. However, pathological biochemical cues or mechanical stress drive VICs toward a myofibroblast phenotype. While this transformation is essential for short-term repair, its persistence contributes to maladaptive MV remodeling and fibrosis. Diabetic stressors may be mitigated …

Mitral Annular Calcification In Diabetes: Valvular Interstitial Cells, Antioxidant Therapy, And Tissue Engineering Approaches, Raleigh J. Keenan

All Theses

Mitral annular calcification (MAC) is a chronic, progressive, and degenerative condition that impairs mitral valve function and increases the risk of cardiovascular disease. The mitral annulus is a saddle-shaped, fibrous structure that is primarily composed of collagen and elastin. The annulus also contains interspersed lipids that are involved in cellular metabolism, as well as inflammatory and osteogenic signaling. These lipids increase the annulus’ susceptibility to calcification, under conditions that increase mechanical stress (hypertension) and promote atherosclerosis (diabetes mellitus). These conditions intensify calcification by promoting the activation of valvular interstitial cells (VICs), increasing fibrosis, and inducing osteogenic differentiation. Although it is …

Light-Induced Phase Separation Purification For Monoclonal Antibodies, Xuyang Chen

All Theses

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Tandem Peptides Targeting Her2 For Delivery Of Cd44 Sirna Into Her2+ Breast Cancer Cells In Vitro, James Kalogeros

All Theses

More than 300,000 women will be diagnosed with breast cancer, and approximately 48,000 women will die this year due to the disease in the United States. HER2+ breast cancer accounts for 20-25% of invasive breast cancer, and it has a worse prognosis and higher early-stage mortality rate than other breast cancer subtypes. Common treatments for HER2+ breast cancer involve surgical removal of the breast or systemic therapy with chemotherapeutic drugs, both of which cause damage to healthy tissue. Due to the lack of a natural ligand for the HER2 receptor, there is a need for improved targeting strategies for the …

Optimizing Crispr/Cas9-Mediated Knockdown Of Angptl3 In Liver Cell Lines And Mouse Hepatocytes, Meredith Reeves

All Theses

Familial hypercholesterolemia (FH) is a genetic condition characterized by elevated levels of low-density lipoprotein cholesterol (LDL-C) that leads to an increased risk of developing cardiac disease early in life (Shah et al., 2020). Current treatments such as statins and PCSK9 inhibitors have helped lower LDL-C levels, however they require repeated administration every 4-6 weeks to remain effective (Raal et al., 2018). Angiopoietin-like 3 (ANGPTL3) is an inhibitor of plasma lipid metabolism that has become a promising molecular target for the treatment of FH. Individuals with non-functional copies of ANGPTL3 demonstrate low levels of plasma LDL-C and triglycerides, indicating a protective …

A 3d Tissue Engineering Model To Study Mitral Valve Annulus Calcification Under Diabetic Conditions, Erin James

All Theses

The most complex heart valve is the mitral valve (MV). Many pathologies can affect the MV, including stenosis, regurgitation, prolapse, and mitral annulus calcification (MAC). MAC is chronic degeneration of the annulus, which is the fibrous, saddle-shaped “ring” that can contract and relax with the myocardium. The prevalence of MAC is around 15% but increases in patients with other cardiovascular diseases and risk factors. It is also thought to increase in patients with type 2 diabetes, but MAC has not been properly characterized within this population because of confounding factors such as cardiac disease and kidney disease. The goal of …

Design Of A Four Channel Pulsatile Perfusion Bioreactor For Ex-Vivo Study Of Vascular Grafts, Thomas M. Fair

All Theses

Cardiovascular diseases are the leading cause of death in the United States. Atherosclerosis in peripheral arteries is a major contributing factor. Autologous saphenous vein grafts are the most common bypass grafts for treating peripheral artery diseases. A quarter of these grafts fail within a year, and around half have failed within 10 years of the initial surgery. Graft failure is attributed to the development of intimal hyperplasia indicated by the migration and proliferation of vascular smooth muscle cells, fibroblasts, and the deposition of extracellular proteins.

Increases in flow, pressure, the pulse frequency, and the differential of pressure that model an …

Investigation On Effects Of Lactate Adaptation Using Metabolic Indicators And Amino Acids To Explore Cell Metabolism, Lisa Uy

All Theses

Biopharmaceutical products are drugs that are made from living cells. One protein type of high importance to biopharmaceuticals is monoclonal antibodies. Monoclonal antibodies allow for highly specific, targeted treatment of diseases and detection of molecules related to diseases. With major roles in detecting and/or treating diseases like human immunodeficiency virus (HIV), coronavirus disease 2019 (COVID-19), and some cancers, monoclonal antibodies are in demand. This demand has led to a need for efficient cell culture systems to produce high yields of protein with minimal waste. The most commonly used cell culture system for these high-value proteins is Chinese hamster ovary (CHO) …