Abstract
The demand for meat is expected to exceed production capacity by livestock in the coming decennia. Therefore, cultured beef might be a viable alternative to traditional livestock-derived beef. One of the problems however is the sustainability of cultured beef through the use of fetal bovine serum. We aimed to identify a serum-free medium or a serum-replacement that is as effective as the current method used for culturing bovine myoblasts. Cells were harvested from a female Blanc Bleu Belge cow and myoblasts were subsequently isolated. Cells were cultured in either Advanced DMEM containing 20% FBS and 10% HS or one of the chemically-defined, serum-free media for 6 days. MTS was used as a measure of cell proliferation at day 1, 4 or 6 and microscopic pictures were taken to assess cell morphology. FBM™, TesR™ and Essential 8™ are commercially available xeno-free media developed for human PSCs and fibroblasts, with the highest potential to sustain bovine myoblast proliferation. Of the supplements tested, XenoFree™ and a custom-prepared growth factor mix failed to stimulate cell proliferation. LipoGro™ stimulated cell proliferation in some cases but also changed the phenotype of myoblasts to an adipocyte-like phenotype. We conclude that serum-free media stimulate exponential cell expansion, albeit not to the extent of the current growth medium containing up to 30% serum. Further research is needed to investigate whether prolonged cell culture or an adaptation period could further increase cell proliferation.
Highlights
Large scale production of primary mammalian cells is crucial to the success of cell therapy and tissue engineering for medical applications, but even more so for upcoming biotechnological solutions for commodities such as meat and leather
Our results are in direct agreement with Skubis et al study on adipose derived MSCs, where a significant decrease in cell viability and increased mitochondrial oxidative activity was observed after 24 h, 48 h and 72 h of treatment of the cells with PSA (Skubis et al 2017)
As LipogroTM seemed to be a suitable supplement to replace serum in terms of supporting cell proliferation; we further investigated the phenotype of the cells (Fig. 7)
Summary
Large scale production of primary mammalian cells is crucial to the success of cell therapy and tissue engineering for medical applications, but even more so for upcoming biotechnological solutions for commodities such as meat and leather. Production methods should be sustainable and cannot rely on serum or other animal-derived materials that are nonreplicative and of limited supply (Post 2014). The use of serum is undesirable from animal welfare and regulatory perspectives. From a financial point of view, the FBS market is very dynamic, leading to continuous price fluctuations rendering it unsustainable for large scale production. Price volatility may not be a major issue in supplies for biomedical research, but will be devastating for large scale cell production to produce for example, meat
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