Serum-free medium suitable for human myoblast culture

doi:10.3969/j.issn.2095-4344.2015.51.013

Abstract

Abstract:

BACKGROUND: Extensive use of fetal bovine serum (FBS) medium to culture myoblasts in vitro has many disadvantages. To develop a more stable, safe, economical serum-free medium that has broad application prospect is indispensable.
OBJECTIVE: To preliminarily explore the serum-free medium suitable for the cultivation of human myoblast.
METHODS: Three different culture media were made: FBS culture medium (DMEM/F12 containing insulin-like growth factor and basic fibroblast growth factor at a ratio of 1:1 and 20% FBS), serum-free medium of mesenchymal stem cells (MSCs; serum-free medium, 2% serum replacement, 2 mmol/L L-glutamine, human umbilical cord-derived MSCs with a purity of > 99%, UltraCULTURETM containing serum replacement and glutamine), and self-made serum-free medium (GibcoTM containing insulin-like growth factor-1, basic fibroblast growth factor, and glutamine). These three media were used to culture human myoblasts respectively. Human
myoblasts were observed morphologically and identified immunohistochemically. Colony forming efficiency was calculated, growth curve was drawn, and viability of human myoblasts was detected using MTT test. Proliferation and differentiation of human myoblasts cultured in the three media were compared and analyzed.
RESULTS AND CONCLUSION: There was no obvious difference in the morphology of the cells among different groups. The purification rate of human myoblasts was up to 99% in each group. The colony forming efficiency of FBS group and self-made serum-free medium group was significantly higher than that of MSCs group (P < 0.01), and the colony forming efficiency of FBS group was significantly higher than that of self-made serum-free medium group (P < 0.01). The viable cell count in the FBS group and self-made serum-free medium group were much higher than that of MSCs group. The cell activity of self-made serum-free medium group was significantly higher than that of FBS group and MSCs group (P < 0.01), while there was no significant difference between FBS group and MSCs group (P > 0.05). Self-made serum-free medium is effective for the culture of myoblasts, and further studies on the early adhesion and proliferation of human myoblasts are necessary.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

CLC Number:

R318

Wang Lin, Liu Mou-yuan, Zhou Li-dong, Liu Li-jin, Liu Ai-bing. Serum-free medium suitable for human myoblast culture[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(51): 8271-8275.

Figures/Tables 5

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