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Title: Characterisation of primary porcine hepatocyte culture systems for use in bioartificial liver devices
Author: Nelson, Leonard Joseph
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2002
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The principal aims of this thesis were to develop a high viability, high yield, reproducible ex vivo method for obtaining PPHs and to characterise, using functional and morphological criteria, PPHs under different fully-defined culture conditions, for use in BAL devices. High yield PPHs isolations, with high viability and purity, cultured under separate SFCD formulations, allow maintainance of viable hepatocyte cultures following high plating efficiencies, compatible with retention of functional activity, for up to 6 days. Moreover, the type of medium formulation appears to modulate, to varying degrees, both cell culture morphology and functional activity in vitro. USR, ASR, tP450, and GE functions for day 2 test media cultures, were up to 300%, 90%, 20%, and 50% respectively, of in vivo values for human liver. Medium 199 is the most suitable synthetic media for culturing PPHs in BALS, which require rapid attachment of cells with an in vivo-like morphology, whilst maintaining a diverse repertoire of hepatic support functions. Furthermore, compared with 2D static cultures, the RCCS modality promotes culture longevity, and cellular functionality (on a per viable cell basis), while the presence of bile canalicular-like structures, and integrin α5expression, suggest the viable cell population maintained a differentiated, 3D-hepatospheroid cytoarchitecture with evidence of polarity. Further study is required to enhance the viable cell population, phenotype, polarity and thus functional activity of such liver constructs, for future therapeutic use.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available