Define of Optimal Addition Period of Osteogenic Peptide to Accelerate the Osteogenic Differentiation of Human Pluripotent Stem Cells

被引:2
作者
Song, Yameng [1 ]
Li, Hongjiao [1 ]
Wang, Zixuan [1 ]
Shi, Jiamin [2 ]
Li, Jing [1 ]
Wang, Lu [1 ]
Liao, Lingzi [1 ]
Ma, Shengqin [1 ]
Zhang, Yun [3 ]
Liu, Bin [1 ]
Yang, Yaling [3 ]
Zhou, Ping [1 ,4 ]
机构
[1] Lanzhou Univ, Sch & Hosp Stomatol, Lanzhou 730000, Peoples R China
[2] Lanzhou Univ, Sch Life Sci, Lanzhou 730000, Peoples R China
[3] Lanzhou Hosp Stomatol, Lanzhou 730000, Peoples R China
[4] Lanzhou Univ, Dept Orthoped, Hosp 2, 82 Cuiyingmen St, Lanzhou 730030, Gansu, Peoples R China
基金
中国博士后科学基金;
关键词
Human pluripotent stem cells; Osteogenic differentiation; Bone forming peptide; Supplement period; Epithelial-mesenchymal transition; BONE MORPHOGENETIC PROTEINS; IN-VITRO; REGENERATION; OSTEOPONTIN; SCAFFOLDS; DELIVERY; RHBMP-2; RELEASE;
D O I
10.1007/s13770-023-00597-y
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Background:The addition of growth factiors is commonly applied to improve the osteogenic differentiation of stem cells. However, for human pluripotent stem cells (hPSCs), their complex differentiation processes result in the unknown effect at different stages. In this study, we focused on the widely used bone forming peptide-1 (BFP-1) and investigated the effect and mechanisms of its addition on the osteogenic induction of hPSCs as a function of the supplementation period.Methods:Monolayer-cultured hPSCs were cultured in osteogenic induction medium for 28 days, and the effect of BFP-1 peptide addition at varying weeks was examined. After differentiation for varying days (0, 7, 14, 21 and 28), the differentiation efficiency was determined by RT-PCR, flow cytometry, immunofluorescence, and alizarin red staining assays. Moreover, the expression of marker genes related to germ layers and epithelial-mesenchymal transition (EMT) was investigated at day 7.Results:Peptide treatment during the first week promoted the generation of mesoderm cells and mesenchymal-like cells from hiPSCs. Then, the upregulated expression of osteogenesis marker genes/proteins was detected in both hESCs and hiPSCs during subsequent inductions with BFP-1 peptide treatment. Fortunately, further experimental design confirmed that treating the BFP-1 peptide during 7-21 days showed even better performance for hESCs but was ineffective for hiPSCs.Conclusion:The differentiation efficiency of cells could be improved by determining the optimal treatment period. Our study has great value in maximizing the differentiation of hPSCs by adding osteogenesis peptides based on the revealed mechanisms and promoting the application of hPSCs in bone tissue regeneration.
引用
收藏
页码:291 / 308
页数:18
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