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Standardizing Enteroid Culture for I...

Klett, Katarina Christina,

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Standardizing Enteroid Culture for Intestinal Radiotherapy Modeling /

Record Type:	Electronic resources : Monograph/item
Title/Author:	Standardizing Enteroid Culture for Intestinal Radiotherapy Modeling // Katarina Christina Klett.
Author:	Klett, Katarina Christina,
Description:	1 electronic resource (160 pages)
Notes:	Source: Dissertations Abstracts International, Volume: 85-06, Section: B.
Contained By:	Dissertations Abstracts International85-06B.
Subject:	Gastrointestinal diseases. -
Online resource:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=30726928
ISBN:	9798381020595

Standardizing Enteroid Culture for Intestinal Radiotherapy Modeling /
Klett, Katarina Christina,

Standardizing Enteroid Culture for Intestinal Radiotherapy Modeling /Katarina Christina Klett. - 1 electronic resource (160 pages)

Source: Dissertations Abstracts International, Volume: 85-06, Section: B.

Enteroids are self-organizing, three-dimensional (3D) cell clusters that mimic native intestinal tissue and offer an alternative to traditional two-dimensional (2D) and in vivomodels. Enteroids contain a variety of intestinal cell types and mimic tissue architecture. While similarly expandable and affordable to traditional 2D cell culture, enteroids better recapitulate the 3D microenvironment. Enteroids can also be derived from patient-biopsies, providing a human-specific alternative to animal models. The current standard of culturing enteroids is within basement membrane extract (BME) that is derived from the matrices surrounding mouse sarcoma. Although enteroids form and grow efficiently within BME matrices, these matrices are inherently variable and cannot be easily tuned. In the second chapter of this thesis, we present a defined, engineered matrix that overcomes these limitations. Specifically, we pair Hyaluronan (HA) with Elastin-Like Protein (ELP) to form the HELP matrix. The HELP matrix supports the formation, growth, and long-term culture of primary murine and human enteroids. We demonstrate the independent tuning of the following matrix properties in HELP: 1) ligand concentration (specifically the fibronectin mimicking ligand RGD), 2) stiffness, and 3) viscoelasticity.In the third chapter of this thesis, we leverage the enteroid model to explore the effects of conventional (CONV) and ultra-high dose rate (FLASH) radiation on healthy intestinal tissue. Radiation is especially damaging to intestinal tissue due to its highly proliferative nature. Fortunately, recent work has demonstrated the potential for ultra-high dose rate delivery to effectively ablate cancerous tissue, while mitigating damage to healthy tissue. While promising, much of this work has relied on murine or 2D cell culture models. In this thesis, we demonstrate the radiation of enteroids derived from healthy primary human intestinal tissue and observe cellular and molecular level differences between CONV- and FLASH-irradiated groups. In summary, this thesis describes: an engineered matrix that was designed to standardize enteroid culture (Chapter 2) and the application of enteroids to investigate intestinal response to radiation (Chapter 3).

English

ISBN: 9798381020595Subjects--Topical Terms:

3821809
Gastrointestinal diseases.

Standardizing Enteroid Culture for Intestinal Radiotherapy Modeling /
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520 $a Enteroids are self-organizing, three-dimensional (3D) cell clusters that mimic native intestinal tissue and offer an alternative to traditional two-dimensional (2D) and in vivomodels. Enteroids contain a variety of intestinal cell types and mimic tissue architecture. While similarly expandable and affordable to traditional 2D cell culture, enteroids better recapitulate the 3D microenvironment. Enteroids can also be derived from patient-biopsies, providing a human-specific alternative to animal models. The current standard of culturing enteroids is within basement membrane extract (BME) that is derived from the matrices surrounding mouse sarcoma. Although enteroids form and grow efficiently within BME matrices, these matrices are inherently variable and cannot be easily tuned. In the second chapter of this thesis, we present a defined, engineered matrix that overcomes these limitations. Specifically, we pair Hyaluronan (HA) with Elastin-Like Protein (ELP) to form the HELP matrix. The HELP matrix supports the formation, growth, and long-term culture of primary murine and human enteroids. We demonstrate the independent tuning of the following matrix properties in HELP: 1) ligand concentration (specifically the fibronectin mimicking ligand RGD), 2) stiffness, and 3) viscoelasticity.In the third chapter of this thesis, we leverage the enteroid model to explore the effects of conventional (CONV) and ultra-high dose rate (FLASH) radiation on healthy intestinal tissue. Radiation is especially damaging to intestinal tissue due to its highly proliferative nature. Fortunately, recent work has demonstrated the potential for ultra-high dose rate delivery to effectively ablate cancerous tissue, while mitigating damage to healthy tissue. While promising, much of this work has relied on murine or 2D cell culture models. In this thesis, we demonstrate the radiation of enteroids derived from healthy primary human intestinal tissue and observe cellular and molecular level differences between CONV- and FLASH-irradiated groups. In summary, this thesis describes: an engineered matrix that was designed to standardize enteroid culture (Chapter 2) and the application of enteroids to investigate intestinal response to radiation (Chapter 3).
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