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- W2980397316 abstract "Standard 2D cell culture does not recreate the complex features of the in vivo environment,such as soluble factor gradients, cell migration into multiple planes, or cell-cell and cell-matrixinteractions.1–3 3D cell culture addresses these limitations by using 3D biomaterial scaffolds,such as alginate hydrogels, to recreate the in vivo cell microenvironment in vitro.4, 5 3Dplatforms can be used to create gradients of soluble factors, vary the biomaterial substratestiffness, permit cell-matrix interactions or promote cell migration.6, 7 Currently available3D platforms are prone to the burst release of soluble factors from the biomaterials, makingit difficult to tightly control the soluble factor concentration.7 This limits the use of 3Dplatforms for investigating processes such as patterned neuronal differentiation, or cell fatespecification in response to small changes in soluble factor concentration.This project proposes a novel 3D alginate platform for patterned differentiation. The firstpart of this thesis describes experiments to optimise alginate hydrogels for the encapsulation,aggregation and differentiation of embryonic stem cells (ESCs), and demonstrates that encapsulated ESCs form embryoid bodies containing cells from the three germ layers. Exogenousretinoic acid (RA) is used for in vitro neuronal differentiation protocols, but exogenous RAis not stable in cell culture and is easily degraded by light. The second part of the thesisoutlines experiments to validate a cell-derived source of RA, which produces a stable concentration of RA in vitro and addresses the limitations of exogenous RA. The final sectiondescribes the novel 3D platform that combines the results from the previous sections using anadapted gradient maker protocol, to create 3D co-culture alginate tubes. The tubes supportpatterned differentiation of ESCs in response to the concentration gradient of cell-derivedRA incorporated into the platform.The novel 3D platform produced in this project contributes a novel tool to the field of3D cell culture. The 3D platform is a tool for investigating ESC differentiation in responseto a 3D concentration gradient of a cell-derived source of retinoic acid. For experiments thatrequire a gradient of RA, the ability to maintain a stable source of RA over several days isan advantage of using this 3D platform over the currently available alternatives. In addition,alginate hydrogels are highly tunable. Thus, the ability to tune the scaffold properties, changethe cell types encapsulated, or introduce gradients of alternative soluble factors makes this aversatile tool for 3D culture." @default.
- W2980397316 created "2019-10-25" @default.
- W2980397316 creator A5043125718 @default.
- W2980397316 date "2018-01-01" @default.
- W2980397316 modified "2023-09-23" @default.
- W2980397316 title "Developing a novel 3D alginate platform for investigating the patterned differentiation of mouse embryonic stem cells" @default.
- W2980397316 hasPublicationYear "2018" @default.
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