Zucchelli, Eleonora;
(2019)
In vivo and in vitro characterization of microtic ear cartilage: potential of patient-derived ear and fat stem cells for cartilage engineering.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
Cartilage has limited regenerative capability, hence its repair in patients with congenital malformations, or following trauma, poses a major challenge for reconstructive surgery. Currently, the most successful ear reconstruction in children affected by microtia (small, abnormally shaped ears) is achieved with autologous tissue grafts carved from rib cartilage requiring several surgeries. Developing autologous tissue-engineered cartilage will reduce surgeries and morbidity at the donor site. My project aimed to assess growth and differentiation potential of progenitors isolated from remnant cartilage of microtic ears in vitro, and compare them to those of rib and adipose-derived stem cells (ADSC), to identify the most suitable cell source(s) for autologous ear reconstruction in paediatric patients affected by microtia. First, microtic and normal ear cartilages were compared, to establish a reference for assessing tissue engineered in vitro. As shown by histology, structural organisation of cartilage was largely lost in microtic ears, although chondrocytes appeared terminally differentiated. Matrisome (extracellular matrix, ECM, protein composition) analysis by proteomics and immunohistochemistry, identified differences between microtic and normal ear cartilage, such as high amount of Collagen-1 and Vimentin, normally not expressed in cartilage at significant levels. Second, chondrogenic differentiation was compared in patients’ normal and microtic ear/rib-derived precursors and ADSCs, grown either in 2-dimensional (2D) or 3D cultures. The differentiation was more efficient in ear-derived cultures, and structural self-organisation was observed in chondrogenic ear-, but not ADSC-, derived spheroids, which resembles the tissue of origin (normal or microtic cartilage) of progenitor cells. In order to assess chondrogenesis in whole-mount spheroids, a new clearing protocol for cartilage was successfully developed. Together, results in 2D and 3D cultures suggest that microtic ear-derived cells can be a suitable cell source for autologous cell therapy. The cartilage matrisome will provide a useful tool for assessing microtic ear chondroblast full maturation potential, for functional cartilage reconstruction.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | In vivo and in vitro characterization of microtic ear cartilage: potential of patient-derived ear and fat stem cells for cartilage engineering |
| Event: | UCL (University College London) |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2019. Original content in this thesis is licensed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) Licence (https://creativecommons.org/licenses/by/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request. |
| UCL classification: | UCL |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10066518 |
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