Orthopaedic tissue engineering utilising immuno-selected human mesenchymal cells
Orthopaedic tissue engineering utilising immuno-selected human mesenchymal cells
In this study a mesenchymal cell population was isolated using the antibody STRO-1 and tested for the ability to generate bone and cartilage. The chondrogenic ability of the isolated cells was tested by generation of aggrecan and type II collagen using micromass culture. For osteogenic tissue generation, selected mesenchymal stem cells were modified to produce BMP-2 and cultured on PLA scaffolds within in vivo diffusion chambers. For alternative tissue engineering strategies, PGA fleece, Porifera, supercritical CO2 formed PLA and alginate cell supports were used as growth conduits. Growth conditions were optimised by using different seeding strategies, combined with perfused and rotating culture vessels to enhance nutrient exchange.
The selection of STRO-1+ cells resulted in a population consisting of 7%±3% of the nucleated marrow cells. In culture the STRO-1+ CFU-F colonies were larger in diameter and were able to express more alkaline phosphatase activity than unselected control cells. Using an in vivo diffusion chamber assay, STRO-1 mesenchymal stem cells formed mineralised, radio-opaque, alkaline phosphatase active, organised type I collagen material within the biodegradable PLA scaffold. In rotating culture with PGA fleece, the continuous cell lines ATDC5 and pZIP were able to produce coherent tissue-engineered constructs, thereby proving the viability of the rotating culture system. Furthermore, STRO-1+ mesenchymal stem cells could be grown throughout PGA fleece scaffolds in rotating and perfused culture. However, culture conditions will have to be optimised for the use of aged primary human cells.
In conclusion, the methods used were able to support a scheme of cartilage and bone tissue engineering and to enhance progenitor cell numbers in vitro, although modification and integration of the constructs will have to be further investigated.
University of Southampton
Howard, Daniel
40caa4c6-e523-4674-8973-992f2b1c5583
2003
Howard, Daniel
40caa4c6-e523-4674-8973-992f2b1c5583
Howard, Daniel
(2003)
Orthopaedic tissue engineering utilising immuno-selected human mesenchymal cells.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
In this study a mesenchymal cell population was isolated using the antibody STRO-1 and tested for the ability to generate bone and cartilage. The chondrogenic ability of the isolated cells was tested by generation of aggrecan and type II collagen using micromass culture. For osteogenic tissue generation, selected mesenchymal stem cells were modified to produce BMP-2 and cultured on PLA scaffolds within in vivo diffusion chambers. For alternative tissue engineering strategies, PGA fleece, Porifera, supercritical CO2 formed PLA and alginate cell supports were used as growth conduits. Growth conditions were optimised by using different seeding strategies, combined with perfused and rotating culture vessels to enhance nutrient exchange.
The selection of STRO-1+ cells resulted in a population consisting of 7%±3% of the nucleated marrow cells. In culture the STRO-1+ CFU-F colonies were larger in diameter and were able to express more alkaline phosphatase activity than unselected control cells. Using an in vivo diffusion chamber assay, STRO-1 mesenchymal stem cells formed mineralised, radio-opaque, alkaline phosphatase active, organised type I collagen material within the biodegradable PLA scaffold. In rotating culture with PGA fleece, the continuous cell lines ATDC5 and pZIP were able to produce coherent tissue-engineered constructs, thereby proving the viability of the rotating culture system. Furthermore, STRO-1+ mesenchymal stem cells could be grown throughout PGA fleece scaffolds in rotating and perfused culture. However, culture conditions will have to be optimised for the use of aged primary human cells.
In conclusion, the methods used were able to support a scheme of cartilage and bone tissue engineering and to enhance progenitor cell numbers in vitro, although modification and integration of the constructs will have to be further investigated.
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Published date: 2003
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Local EPrints ID: 465171
URI: http://eprints.soton.ac.uk/id/eprint/465171
PURE UUID: 2ccf4d18-e3a5-47c2-a7fa-e868891b2f18
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Date deposited: 05 Jul 2022 00:27
Last modified: 16 Mar 2024 20:00
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Author:
Daniel Howard
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