Accessible Primary Cell Isolation and Culture for Cultured Meat Bioprocessing

: (Alternative Format Thesis)

Abstract

Cellular agriculture aims to recreate and replace products such as milk, meat, and leather with cultivated cells and/or their products to offer a more ethical, sustainable alternative to traditional agriculture. With the goal of addressing a subset of the scientific obstacles facing cultured meat production, the main objectives of this research were (1) to determine whether primary cells may be isolated from commercial meat products; (2) to develop methods to improve cell yields from commercial meat products; (3) to characterise the primary cell populations derived from supermarket chicken breast; and (4) to investigate the suitability of the derived primary chicken cells for incorporation in hollow fibre bioreactors (HFBs). After a meat screening and method development phase, pre-packaged supermarket chicken breast was selected as the highest-yielding source tissue (when compared with butcher’s pork and pre-packaged supermarket lamb, pork, and beef) and an isolation protocol based on explant outgrowth was established and cells derived from explant isolations were cultured and characterised. Chicken explants were then cultured over three outgrowth phases, and cells from each outgrowth phase were cultured for three passages for characterisation. These cells demonstrated doubling times ranging from 33.2-55.0 hours, myogenic purities ranging from ~50-80%, and fusion indices ranging from 0.5-2.5%. Attempts to adapt this isolation protocol to animal component-free conditions failed due to insufficient outgrowth and stagnant cell populations. The longevity of the explant tissues was then explored, and explants exhibited the ability outgrow cells to confluence, inducing cells to detach. The detached cells were then capable of reattachment and continued growth, with a reattachment efficiency of ~60-90%, paving the way for a continuous harvest paradigm, either for sourcing cells from explant dishes or for harvesting continuously-detaching cells from bioreactor scaffolds as confluence is achieved. Finally, the primary chicken cells were tested for their compatibility with PX40, a scaffold material used to make the fibres in HFBs, and achieved attachment efficiencies of ~28-43% after 3 hours, which were no different from attachment on tissue culture plastic. In all, the objectives of the research were achieved, an accessible source of primary chicken cells was established, and the potential of the cells for use in HFB bioprocessing was validated.

Date of Award	25 Jun 2025
Original language	English
Awarding Institution	University of Bath
Sponsors	New Harvest
Supervisor	Marianne Ellis (Supervisor), David Tosh (Supervisor) & Chris Chuck (Supervisor)