Cell Control Via Micropatterning

Abstract

Institution Name: The University of ManchesterCandidate Name: Naa-Dei NikoiDegree Title: Master of Philosophy (MPhil)Thesis Title: Cell Control Via MicropatterningDate: May 2012Cell morphology has a profound effect on cell function, a relationship that is mediated by the effects of surface adhesion on focal adhesions and of geometry on cytoskeletal architecture. The basic principles of mechanotransduction from focal adhesion to the nucleus have been elucidated, but its specific effect on primary cells is still rarely-studied. In this study, primary human osteoblasts have been used as model cells for the investigation of the effect of shape on cell function. Work done focussed on developing a cell restriction method to robustly enclose single cells or small numbers of cells on a two-dimensional surface in order to separate the effect of shape from that of topography. Patterns for restriction were selected on their ability to present different amounts of contractility. Surface area was chosen on the basis of the largest cells seen in unrestricted cells. Three methods of restricting cells to surfaces (PEG-terminated thioalkane on gold, NHS-conjugated PEG and PEG-PPO-PEG tri-block copolymers on glass substrates) were developed in-house. A fourth, PEG-terminated silanes on glass, was developed elsewhere and was also evaluated. Cells were successfully restricted on surfaces for 1-2 days using PEG-thiolalkanes, tri-block copolymers and PEG-terminated silanes. NHS-conjugated PEG surfaces successfully restricted a simple protein solution, but failed to restrict cells. Where restricted, osteoblasts displayed an unexpected behavioural change, with multiple cells clustering on a shape and on top of one another. Similar clustering was not seen on control surfaces. There is a need to improve the robustness of the restriction methods as well as a need to utilise better patterns. Significant work needs to be done to achieve the objective of building a complete picture of a cell once confined to a shape and approaches are outlined in the final section.

Date of Award	31 Dec 2012
Original language	English
Awarding Institution	The University of Manchester
Supervisor	Julie Gough (Supervisor) & Ian Kinloch (Supervisor)