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The Feasibility of Establishing a Morphological Cell Profiling Assay to Assess the Bioactivity of Natural Products

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thesis
posted on 16.11.2021, 20:52 by Taylor, Lauren

Morphological cell profiling (MCP) is an assay which quantifies the morphology of cells and cellular components. Changes in the morphology of cells following compound treatment has been shown to assess the bioactivity, and even propose a target of said compound. It is a powerful assay that can be used to assess novel compounds for drug candidature. However, it is currently not transferable between institutions. Thus, this project evaluated the feasibility of establishing an MCP assay at Victoria University of Wellington - Te Herenga Waka to assess the bioactivity of novel natural products from the natural products laboratory.

First, a new individualistic approach to select compound concentration for MCP was assessed by flow cytometry with six exemplary drugs. Results concluded this flow cytometry approach can realise an ideal concentration for individual compounds in which the compound treatment was bioactive but not cytotoxic. This approach provides more biologically relevant and accurate information about a compound's bioactivity than previous MCP methods.

Second, the feasibility of establishing the assay was assessed by testing the same six drugs through protocol common to MCP assays. Various stains and microscopes were tested for suitability for an MCP; image quantitative software CellProfiler was evaluated for MCP analysis ease; and quality control protocol was attempted. Ultimately, an MCP is currently not feasible at Te Herenga Waka as multiple components to establish the assay are too difficult and time intensive to complete.

Third, discorhabdin E, a pyrroloiminoquinone alkaloid, was isolated from the New Zealand marine sponge, Latrunculia kaakaariki. The compound was tested as if it were a novel natural product compound through the new flow cytometry concentration selection protocol to test its effectivity, and was subsequently assessed by morphological analysis.

History

Copyright Date

17/11/2021

Date of Award

17/11/2021

Publisher

Victoria University of Wellington - Te Herenga Waka

Rights License

Author Retains Copyright

Degree Discipline

Biomedical Science

Degree Grantor

Victoria University of Wellington - Te Herenga Waka

Degree Level

Masters

Degree Name

Master of Biomedical Science

ANZSRC Type Of Activity code

4 EXPERIMENTAL RESEARCH

Victoria University of Wellington Item Type

Awarded Research Masters Thesis

Language

en_NZ

Victoria University of Wellington School

School of Biological Sciences

Advisors

McConnell, Melanie; Keyzers, Robert