Engineering Tumor Models Using Aqueous Biphasic 3D Culture Microtechnology
| Main Author: | |
|---|---|
| Language: | English |
| Published: |
University of Akron / OhioLINK
2017
|
| Subjects: | |
| Online Access: | http://rave.ohiolink.edu/etdc/view?acc_num=akron150470381711759 |
| id |
ndltd-OhioLink-oai-etd.ohiolink.edu-akron150470381711759 |
|---|---|
| record_format |
oai_dc |
| collection |
NDLTD |
| language |
English |
| sources |
NDLTD |
| topic |
Biomedical Engineering Cellular Biology Biology cancer tumor microenvironment spheroids drug discovery aqueous two-phase system triple negative breast cancer |
| spellingShingle |
Biomedical Engineering Cellular Biology Biology cancer tumor microenvironment spheroids drug discovery aqueous two-phase system triple negative breast cancer Ham, Stephanie Lemmo Engineering Tumor Models Using Aqueous Biphasic 3D Culture Microtechnology |
| author |
Ham, Stephanie Lemmo |
| author_facet |
Ham, Stephanie Lemmo |
| author_sort |
Ham, Stephanie Lemmo |
| title |
Engineering Tumor Models Using Aqueous Biphasic 3D Culture Microtechnology |
| title_short |
Engineering Tumor Models Using Aqueous Biphasic 3D Culture Microtechnology |
| title_full |
Engineering Tumor Models Using Aqueous Biphasic 3D Culture Microtechnology |
| title_fullStr |
Engineering Tumor Models Using Aqueous Biphasic 3D Culture Microtechnology |
| title_full_unstemmed |
Engineering Tumor Models Using Aqueous Biphasic 3D Culture Microtechnology |
| title_sort |
engineering tumor models using aqueous biphasic 3d culture microtechnology |
| publisher |
University of Akron / OhioLINK |
| publishDate |
2017 |
| url |
http://rave.ohiolink.edu/etdc/view?acc_num=akron150470381711759 |
| work_keys_str_mv |
AT hamstephanielemmo engineeringtumormodelsusingaqueousbiphasic3dculturemicrotechnology |
| _version_ |
1719452637285318656 |
| spelling |
ndltd-OhioLink-oai-etd.ohiolink.edu-akron1504703817117592021-08-03T07:04:08Z Engineering Tumor Models Using Aqueous Biphasic 3D Culture Microtechnology Ham, Stephanie Lemmo Biomedical Engineering Cellular Biology Biology cancer tumor microenvironment spheroids drug discovery aqueous two-phase system triple negative breast cancer Cancer remains a leading cause of death in the United States with 1.7 million new cases diagnosed and over half a million deaths reported in 2016 according to the National Cancer Institute. The disease was traditionally considered as a group of cells that become malignant from accumulated mutations. However, this definition is now recognized as being too simplistic. Tumors are comprised of multiple cell types, matrix proteins, and soluble factors and in essence resemble an organ. The microenvironment surrounding the mass of cancer cells constitutes the tumor stroma. Signaling between cancer cells and tumor stroma components such as fibroblast cells, immune cells, vascular cells, and extracellular matrix proteins mediate functional regulation of cancer cells including growth, proliferation, migration, and drug response. Despite the significant influence of the tumor stroma on cancer cell behavior, current in vitro and animal models fail to recreate cancer cell - stroma interactions of human tumors. To overcome existing tumor modeling limitations, we developed an automated and high throughput technology for the generation of three-dimensional (3D) mass of cancer cells, known as a spheroid, that enables the incorporation of tumor stromal components to recreate cancer cell - stroma interactions. This technology utilizes an aqueous two-phase system (ATPS) with dextran (DEX) and polyethylene glycol (PEG) polymer phases and allows facile printing of spheroids. Cancer cells of interest, with or without stromal cells, are entrapped in the DEX phase solution when dispensed as a droplet in PEG phase solution and self-aggregate to form a spheroid. The technology addresses challenges of existing spheroid formation methods by using standardized tools, such as liquid handling robotics and microplates, for reproducible and reliable spheroid formation and convenient downstream biochemical and molecular analysis of spheroids. We focused on triple negative breast cancer (TNBC) as a disease model. TNBC is the most lethal subtype of breast cancer suffering from a lack of targeted therapy options and high tumor stromal content, underscoring the strong unmet need to better understand its aggressive biology for the development of effective treatment options. The role of the tumor stroma on TNBC cell growth and drug response is studied through optimization of the spheroid printing technology, biological validation of the resulting TNBC cell spheroids, and incorporation of cancer-associated fibroblasts (CAFs), the most abundant stromal cell, into TNBC cell spheroids. The ATPS spheroid technology results in consistently sized TNBC spheroids of different cell densities in standard 384-microwell plates. Biological validation of TNBC spheroids shows that they reproduce key properties of solid tumors such as extracellular matrix (ECM) protein deposition, proliferation gradients, hypoxia, and cancer stem cell (CSC) markers expression. Our findings show that interaction with CAFs significantly stimulates TNBC cell growth primarily through upregulated activity of major survival kinase pathways. Additionally, the presence of a 3D environment, expression of hypoxia and CSC markers, and cancer cells-CAFs signaling promote chemotherapy drug resistance of TNBC cells. Importantly, targeting tumor microenvironment factors such as hypoxia and CAFs are shown to be successful in sensitizing TNBC cell spheroids to drug treatment, offering a novel approach to improve drug responses of cancer cells by targeting tumor stroma. This work strongly supports the advantages of the 3D tumor technology to model the complex biology of tumors and facilitate the discovery of novel and promising therapeutic approaches to eventually benefit cancer patients. 2017 English text University of Akron / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=akron150470381711759 http://rave.ohiolink.edu/etdc/view?acc_num=akron150470381711759 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws. |