Comparison of Single Cell Volume Between Two-Dimensional and Three-Dimensional Cell Culture

Manning, Joshua

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Overview

Year:: 2021
Contributor:: Manning, Joshua (creator); Morgan, Jeffrey (Advisor); Ip, Blanche (Reader); Toussaint, Kimani (Reader); Brown University. Biology and Medicine: Biomedical Engineering (sponsor)
Genre:: theses
Subject:: Confocal microscopy; Cell Volume; Microtissues; 3D Cell Culture
Extent:: 7, 71 p.

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Description

Abstract:: Recent advancements in the techniques used for developing in vitro three-dimensional cell cultures, such as bioprinting or microtissue formation, has offered a more accurate representation of in vivo environments when compared to two-dimensional monolayers. However, due to the advanced structure of 3D cultures, new analysis techniques are needed to assess potential physiological variations between individual cells within these constructs. Cell volume regulation has been shown to play a pivotal role in determining cell functionality both in vivo and in vitro. Cellular proliferation and apoptosis are both mechanisms regulated by cell volume. It is necessary to develop a technique for assessing cellular interactions, physiology, and morphology within the inner regions of microtissues. The work of this thesis aims to establish an imaging technique capable of analyzing single cells within 3D microtissue environments, in order to quantify and compare single cell volume between 2D and 3D cultures. Human ovarian granulosa-like tumor cells (KGNs) were seeded within a 3D spheroidal environment using a non-adhesive agarose mold and stained at a 10% ratio using cell tracker green. Using confocal microscopy, individual cells could be distinguished within the greater microtissue construct and analyzed for cell volume. The volume of single cells within the 3D microtissue appeared to decrease when compared to cells within 2D culture environments. In addition, cells located at the interior positions of the spheroid showed lower volumes when compared to cells located at the outer regions. The results of this thesis aims to provide a high-throughput technique for observing individual cells within a microtissue construct. Having a high-throughput method will allow for assessment of physiological variations of individual cells within 3D cell culture environments.
Notes:: Thesis (Sc. M.)--Brown University, 2021

Content

Citation

Manning, Joshua, "Comparison of Single Cell Volume Between Two-Dimensional and Three-Dimensional Cell Culture" (2021). Biomedical Engineering Theses and Dissertations. Brown Digital Repository. Brown University Library. https://repository.library.brown.edu/studio/item/bdr:qxhg7asr/

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Biomedical Engineering Theses and Dissertations

Theses and Dissertations for the Biomedical Engineering department.
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