Bovine Satellite Cells Isolated after 2 and 5 Days of Tissue Storage Maintain the Proliferative and Myogenic Capacity Needed for Cultured Meat Production

Abstract

Cultured meat is an emerging alternative food technology which aims to deliver a more ethical, sustainable, and healthy muscle-tissue-derived food item compared to conventional meat. As start-up companies are rapidly forming and accelerating this technology, many aspects of this multi-faceted science have still not been investigated in academia. In this study, we investigated if bovine satellite cells with the ability to proliferate and undergo myogenic differentiation could be isolated after extended tissue storage, for the purpose of increasing the practicality for cultured meat production. Proliferation of bovine satellite cells isolated on the day of arrival or after 2 and 5 days of tissue storage were analyzed by metabolic and DNA-based assays, while their myogenic characteristics were investigated using RT-qPCR and immunofluorescence. Extended tissue storage up to 5 days did not negatively affect proliferation nor the ability to undergo fusion and create myosin heavy chain-positive myotubes. The expression patterns of myogenic and muscle-specific genes were also not affected after tissue storage. In fact, the data indicated a positive trend in terms of myogenic potential after tissue storage, although it was non-significant. These results suggest that the timeframe of which viable myogenic satellite cells can be isolated and used for cultured meat production can be greatly extended by proper tissue storage.

Recombinant production of growth factors for application in cell culture

“…To assess the suitability of the recombinantly-produced GF for use in cultured meat applications we assayed for bioactivity on BSC. The BSC used were isolated from three different donor animals and have been previously shown to express myogenic markers and to form myosin heavy chain-positive myotubes ( Skrivergaard et al., 2021 ). Using these cells, we screened multiple orthologs from each GF family in seven different concentrations ranging from 1.56 to 100 ng/mL.…”

“…Satellite cells were isolated from three different bovine donors as described in ( Skrivergaard et al., 2021 ). The donors (3–4 yearold Holstein dairy cows) were transported to a commercial plant (Danish Crown, Aalborg, Denmark), and slaughtered within 1 h after arrival by stunning with captive bolt followed by bleeding and electrical stunning at 100 V for 400 s. The M usculus semitendinosus was removed within 20 min postmortem and transported on ice to the cell laboratory at department of Food Science, Aarhus University.…”

“…Satellite cells that were previously isolated from three different bovine donors ( Skrivergaard et al., 2021 ) were expanded in Matrigel coated T75 flasks (Nunc cat. no.…”

Review of the Current Research on Fetal Bovine Serum and the Development of Cultured Meat

“…The acquisition of raw cells is essential for producing cultured meat, and it is very important to control cell stability parameters. Skrivergaard et al (2021) studied cell proliferation and differentiation according to the storage period of the bovine muscle tissue for cell acquisition. Satellite cells isolated from tissues stored under refrigeration at 4°C were poorly cultured at the initial cultivation compared to the satellite cells isolated from fresh cultures, but it was found that there was no difference in metabolic activity and DNA concentration.…”

Repeated and long-term cryopreservation of primary bovine myogenic cells to maintain quality in biomimetic cultured meat

“…Long-term storage of cultured cells, as a cell source for cultured meat, is one technique urgently needed to be addressed. Stig Skrivergaard et al demonstrated in their previous study that bovine satellite cells were able to maintain their important properties after muscle tissue storage at 4 • C for up to 5 days (Skrivergaard et al, 2021). Since cryopreservation is a well-established procedure in other research fields, few studies demonstrated that bovine myogenic cells were frozen in studies of cultured meat production (Ding et al, 2018;Kolkmann et al, 2020;Skrivergaard et al, 2021;Andreassen et al, 2022).…”