Davidson, Shanna-Leigh
(2023)
Designing Functional Nanocultures for Controlled Microbial Dynamics.
Doctoral Dissertation, University of Pittsburgh.
(Unpublished)
Abstract
While new techniques and platforms have advanced rapidly to enhance our understanding of complex microbial dynamics utilizing culture-independent methods, the same cannot be said for wet-lab culturing techniques. In fact, developments in meta-omics have revealed large bottlenecks in cell-culturing; omics methods can quickly reveal a myriad of information on microbial populations, yet, recapitulating those same microbial populations in a controlled environment is a non-trivial task. Therefore, there is a critical need to understand how the local milieu of microorganisms facilitates their growth in natural environments. Miniaturized cell-culturing techniques and novel microsystems can facilitate the interrogation of microorganisms and their local milieu in well-defined environments. Innovatively designed micro-technologies have shifted the paradigm in conventional microbial culturing. However, many of these technologies do not provide robust platforms for long-term studies, and are further limited by singular application; therefore, do not translate well from lab to clinical applications. Aspiring to design a novel tool for these purposes, our team has developed Nanocultures: nanolitre-sized, double-emulsion, polymeric microcapsules to sequester and cultivate microbial consortia. The nanocultures provide a robust, optically transparent, and high-throughput tool to study cell dynamics. As semi-permeability is a key design parameter, we can manipulate the chemistry of the nanoculture shell to achieve desired functionalities, such as for high-throughput screening and the development of biotherapeutics to reconstitute a disturbed microflora. As such, this thesis aims to explore I) transport and II) mechanical properties of a newly designed polymer, providing a highly functionalized shell with advantageous semi-permeability properties to the encased bacteria, as well as beneficial mechanical properties that allow for targeted lysis of the polymeric shell. Thirdly, this thesis discusses the design of a novel co-culturing platform to enable the growth of mammalian and bacterial cells together and allows for the assessment of cell-cell interactions in a first attempt, and proof-of-concept design of clinically applicable biotherapeutics. Together, these data will pave the way in our understanding of designing optimal nanocultures for applications-based functionality.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
|
| ETD Committee: |
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| Date: |
14 September 2023 |
| Date Type: |
Publication |
| Defense Date: |
5 May 2023 |
| Approval Date: |
14 September 2023 |
| Submission Date: |
24 July 2023 |
| Access Restriction: |
2 year -- Restrict access to University of Pittsburgh for a period of 2 years. |
| Number of Pages: |
175 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
Swanson School of Engineering > Chemical and Petroleum Engineering |
| Degree: |
PhD - Doctor of Philosophy |
| Thesis Type: |
Doctoral Dissertation |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
micro-culturing, micro-technology, droplet Microfluidics, double-emulsion, Flory-Huggins, interaction parameter, small molecule diffusion, mechanical disruption, multimodal -omics platform, co-culture, Caco-2 |
| Date Deposited: |
14 Sep 2023 13:43 |
| Last Modified: |
14 Sep 2023 13:43 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/45140 |
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