Kolkmann A M, Post M J, Rutjens M A M, van Essen A L M, Moutsatsou P
Dept. Physiology, Maastricht University, 6229 ER, Maastricht, The Netherlands.
Cytotechnology. 2020 Feb;72(1):111-120. doi: 10.1007/s10616-019-00361-y. Epub 2019 Dec 28.
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.
预计在未来十年，肉类需求将超过牲畜的生产能力。因此，养殖牛肉可能是传统畜牧来源牛肉的可行替代品。然而，问题之一是使用胎牛血清培养牛肉的可持续性。我们旨在确定一种无血清培养基或血清替代物，其效果与目前用于培养牛成肌细胞的方法相同。从一头雌性白蓝比利时奶牛身上采集细胞，随后分离出成肌细胞。将细胞培养在含有20%胎牛血清和10%马血清的Advanced DMEM或一种化学成分明确的无血清培养基中6天。在第1、4或6天使用MTS作为细胞增殖的指标，并拍摄显微镜照片以评估细胞形态。FBM™、TesR™和Essential 8™是为人类多能干细胞和成纤维细胞开发的市售无异种培养基，具有维持牛成肌细胞增殖的最大潜力。在测试的补充剂中，XenoFree™和定制的生长因子混合物未能刺激细胞增殖。LipoGro™在某些情况下刺激细胞增殖，但也将成肌细胞的表型改变为脂肪细胞样表型。我们得出结论，无血清培养基可刺激细胞呈指数级扩增，尽管其程度不如目前含高达30%血清的生长培养基。需要进一步研究以调查延长细胞培养时间或适应期是否能进一步增加细胞增殖。
