E-Thesis 331 views 132 downloads
Identification and Characterisation of tracheal cartilage derived stem cells for airway tissue engineering / Navid Moshkbouymatin
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DOI (Published version): 10.23889/Suthesis.53568
The trachea is a complex organ composed of multiple cell types and is just one integral part of the respiratory system and as a result of injury or insult there is an immediate need to engineer a neo-trachea as there currently no long-term treatments for tracheal defects. This thesis has for the fir...
|Supervisor:||Khan, Ilyas ; Francis, Lewis|
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The trachea is a complex organ composed of multiple cell types and is just one integral part of the respiratory system and as a result of injury or insult there is an immediate need to engineer a neo-trachea as there currently no long-term treatments for tracheal defects. This thesis has for the first time successfully identified a mesenchymal derived stem/progenitor cell component that reside within the tracheal C-ring cartilage by means of selective adhesion protocol of fibronectin for airway tissue engineering applications. These cells were found to be, plastic adherent, colony forming, expressed the minimal cell surface markers and were capable of undergoing tri-lineage. Further analysis revealed mechanical property changes in that the tracheal colony forming cells became stiffer with each passage and the gene expression of collagen I, II and X were reduced. To investigate the chondrogenic potential of tracheal stem/progenitor cells traditional pellet culture over a 21-day time course resulted in matrix formation consisting of collagen type II, aggrecan, collagen type I and possible calcification. To rule out the influence of plastic culture 3D gelatin- derived microcarrier culture techniques in both static and wave culture technology were employed for large expansion of tracheal chondroprogenitors. Post expansion analysis revealed that lubricin (PRG4) transcription was observed in all wave expanded cells which is indicative of superficial zone of articular cartilage and not tracheal cartilage. Post differentiation analysis of the microcarrier constructs revealed similar gene profiles to that observed in traditional pellet culture, although with a slight reduction in gene expression. However, microcarrier based differentiation reduced collagen type X gene expression when compared to traditional pellet culture in all the microcarrier groups. These findings taken together show great potential in the wider cartilage research community in that microcarrier and bioreactor expansion can induce the transcription of PRG4 which is specific to the superficial zone of articular cartilage. Furthermore, as a proof concept C-ring like structures were fabricated using tracheal cartilage derived stem/progenitor cells and microcarriers in 3D printed moulds for use as a customisable new method for airway tissue engineering.
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