A dissertation on deterministic culturing of single cells in 3D

Reason: Pending publications and patents.

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Deterministic Culturing of Single Cells in 3D

thesis
posted on 01.03.2021, 23:04 by Rohil Jain
Models using 3D cell culture techniques are increasingly accepted as the most biofidelic in vitro representations of tissues for research. These models are generated using biomatrices and bulk populations of cells derived from tissues or cell lines. This thesis study focuses on an alternate method to culture individually selected cells in relative isolation from the rest of the population under physiologically relevant matrix conditions. Matrix gel islands are spotted on a cell culture dish to act as support for receiving and culturing individual single cells; a glass capillary-based microfluidic setup is used to extract each desired single cell from a population and seed it on top of an island. Using examples of breast and colorectal cancers, we show that individual cells evolve into tumors or aspects of tumors displaying different characteristics of the initial cancer type and aggressiveness. By implementing a morphometry assay with luminal A breast cancer, we demonstrate the potential of the proposed approach to studying phenotypic heterogeneity. Results reveal that intertumor heterogeneity increases with time in culture and that varying degrees of intratumor heterogeneity may originate from individually seeded cells. Moreover, we observe a positive correlation between fast-growing tumors and the size and heterogeneity of their nuclei.

Funding

A novel, reliable and sensitive optical detection platform for ovarian cancer.

Directorate for Engineering

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Tom Hurvis and the McKinley Educational Foundation

History

Degree Type

Doctor of Philosophy

Department

Mechanical Engineering

Campus location

West Lafayette

Advisor/Supervisor/Committee Chair

Cagri Savran

Additional Committee Member 2

Chun-Li Chang

Additional Committee Member 3

Sophie Lelievre

Additional Committee Member 4

Bumsoo Han

Additional Committee Member 5

Babak Ziaie

Licence

Exports