No Cover Image

E-Thesis 517 views 188 downloads

Identification and Characterisation of tracheal cartilage derived stem cells for airway tissue engineering / Navid Moshkbouymatin

DOI (Published version): 10.23889/Suthesis.53568

Abstract

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...

Full description

Published: Swansea 2019
Institution: Swansea University
Degree level: Doctoral
Degree name: Ph.D
Supervisor: Khan, Ilyas ; Francis, Lewis
URI: https://cronfa.swan.ac.uk/Record/cronfa53568
Tags: Add Tag
No Tags, Be the first to tag this record!
first_indexed 2020-02-20T19:59:26Z
last_indexed 2020-09-17T03:16:53Z
id cronfa53568
recordtype RisThesis
fullrecord <?xml version="1.0"?><rfc1807><datestamp>2020-02-18T10:40:25.3002597</datestamp><bib-version>v2</bib-version><id>53568</id><entry>2020-02-18</entry><title>Identification and Characterisation of tracheal cartilage derived stem cells for airway tissue engineering</title><swanseaauthors/><date>2020-02-18</date><abstract>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.</abstract><type>E-Thesis</type><journal/><publisher/><placeOfPublication>Swansea</placeOfPublication><keywords/><publishedDay>20</publishedDay><publishedMonth>8</publishedMonth><publishedYear>2019</publishedYear><publishedDate>2019-08-20</publishedDate><doi>10.23889/Suthesis.53568</doi><url/><notes>A selection of third party content is redacted or is partially redacted from this thesis.</notes><college>COLLEGE NANME</college><CollegeCode>COLLEGE CODE</CollegeCode><institution>Swansea University</institution><supervisor>Khan, Ilyas ; Francis, Lewis</supervisor><degreelevel>Doctoral</degreelevel><degreename>Ph.D</degreename><apcterm/><lastEdited>2020-02-18T10:40:25.3002597</lastEdited><Created>2020-02-18T10:40:25.3002597</Created><authors><author><firstname>Navid</firstname><surname>Moshkbouymatin</surname><order>1</order></author></authors><documents><document><filename>53568__16650__c4ff595286924b48ae669c36016e0773.pdf</filename><originalFilename>Moshkbouymatin_Navid_PhD_Thesis_Final_Embargoed_20.08.2021_Redacted.pdf</originalFilename><uploaded>2020-02-20T15:44:18.4601944</uploaded><type>Output</type><contentLength>8228534</contentLength><contentType>application/pdf</contentType><version>Redacted version - open access</version><cronfaStatus>true</cronfaStatus><embargoDate>2021-08-20T00:00:00.0000000</embargoDate><copyrightCorrect>true</copyrightCorrect></document></documents><OutputDurs/></rfc1807>
spelling 2020-02-18T10:40:25.3002597 v2 53568 2020-02-18 Identification and Characterisation of tracheal cartilage derived stem cells for airway tissue engineering 2020-02-18 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. E-Thesis Swansea 20 8 2019 2019-08-20 10.23889/Suthesis.53568 A selection of third party content is redacted or is partially redacted from this thesis. COLLEGE NANME COLLEGE CODE Swansea University Khan, Ilyas ; Francis, Lewis Doctoral Ph.D 2020-02-18T10:40:25.3002597 2020-02-18T10:40:25.3002597 Navid Moshkbouymatin 1 53568__16650__c4ff595286924b48ae669c36016e0773.pdf Moshkbouymatin_Navid_PhD_Thesis_Final_Embargoed_20.08.2021_Redacted.pdf 2020-02-20T15:44:18.4601944 Output 8228534 application/pdf Redacted version - open access true 2021-08-20T00:00:00.0000000 true
title Identification and Characterisation of tracheal cartilage derived stem cells for airway tissue engineering
spellingShingle Identification and Characterisation of tracheal cartilage derived stem cells for airway tissue engineering
,
title_short Identification and Characterisation of tracheal cartilage derived stem cells for airway tissue engineering
title_full Identification and Characterisation of tracheal cartilage derived stem cells for airway tissue engineering
title_fullStr Identification and Characterisation of tracheal cartilage derived stem cells for airway tissue engineering
title_full_unstemmed Identification and Characterisation of tracheal cartilage derived stem cells for airway tissue engineering
title_sort Identification and Characterisation of tracheal cartilage derived stem cells for airway tissue engineering
author ,
author2 Navid Moshkbouymatin
format E-Thesis
publishDate 2019
institution Swansea University
doi_str_mv 10.23889/Suthesis.53568
document_store_str 1
active_str 0
description 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.
published_date 2019-08-20T04:06:34Z
_version_ 1763753480658354176
score 11.012678