Le cordon ombilical humain, source de cellules pour le génie tissulaire : isolement, caractérisation et production de substituts humains
|Authors:||Hayward, Cindy Jean|
|Advisor:||Auger, François A.; Fradette, Julie|
|Abstract:||The human umbilical cord has received increasing attention as a source of cells for both research and therapeutic purposes. Four main cell types – epithelial, stromal, smooth muscle and endothelial cells – make up the solid tissues of the umbilical cord. Some of these cell types have been used in research for decades, while the potential of others is just being recognised. We have developed a unique protocol for the sequential extraction of all four cell types from a single umbilical cord, thus allowing the reconstruction of tissues and organs with cells from the same source. A combination of perfusion, immersion and explant techniques allows the successful extraction and expansion in culture of these cells. Further characterisation of the epithelial and Wharton’s jelly cells was carried out by immunofluorescent staining of specific proteins. The potential of these cells for use in regenerative medicine was demonstrated through the production of tissue-engineered constructs, including a blood vessel composed of umbilical cord stromal and smooth muscle cells which showed a substantial burst resistance under pressure. The capacity for differentiation of cord epithelial cells was studied in the context of a bilayered reconstructed skin substitute, in combination with keratinocytes, dermal fibroblasts, and Wharton’s jelly cells. These epithelial cells differentiated in a manner similar to keratinocytes when cultured on dermal fibroblasts and exposed to air, but under the same conditions on cord stromal cells they degenerated. Finally, to complete our study the Wharton’s jelly cells were induced to differentiate in vitro into various mesenchymal cell types. Globally, this work shows the importance of not only the culture conditions on the growth and differentiation of the various cell types, but also the important effect of the cell source on the resulting reconstructed tissues.|
|Document Type:||Thèse de doctorat|
|Open Access Date:||24 April 2018|
|Collection:||Thèses et mémoires|
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