Identification of biomarkers indicative of functional skeletal stem cells
Identification of biomarkers indicative of functional skeletal stem cells
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DOI：
10.1111/ocr.12260
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发表时间：
2019-05-01
影响因子：
3.1
通讯作者：
Villa-Diaz, Luis G.
中科院分区：
文献类型：
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作者：
Qiryaqoz, Zeena;Timilsina, Suraj;Villa-Diaz, Luis G.
摘要目的骨干细胞（bone stem cells, SSCs）具有细胞表面生物标志物的表达和向骨、软骨和脂肪分化的能力。然而，目前用于鉴定这些细胞群的生物标志物不是细胞类型特异性的，也不能指示这些细胞的分化状态，因此不可靠。我们的目的是鉴定从骨髓（BM）分离的ssc和来自多能干细胞（PSC）的ssc之间共享的替代细胞表面生物标志物和转录因子。材料与方法将人PSCs诱导成SSCs。使用FACS和qRT-PCR来确定来自PSCs和BM分离的ssc，分化细胞，早期和晚期传代细胞以及成纤维细胞中细胞表面生物标志物和转录因子的表达差异。结果3次传代后，SSCs的增殖能力明显降低，向脂肪细胞和成骨细胞分化的能力明显减弱。蛋白质和mRNA分析表明，常用的生物标志物在失去分化能力的细胞中仍然高度表达。然而，与成纤维细胞相比，整合素- 6 （CD49f）和转录因子GATA6、PRDM16、SIM2和SOX11在ssc中显著上调。在成脂和成骨分化的早期，后期传代细胞中CD49f、GATA6和SIM2的表达降低，增殖和分化能力有限。结论CD49f和转录因子GATA6和SIM2鉴定功能性ssc。
Structured AbstractObjectives Skeletal stem cells (SSCs) are characterized by expression of cell surface biomarkers and their ability to differentiate into bone, cartilage and fat. However, the current biomarkers used to identify these cell populations are not cell-type-specific or indicative of the differentiation status of these cells and are therefore unreliable. Our objective was to identify alternative cell surface biomarkers and transcription factors shared between SSCs isolated from the bone marrow (BM) and those derived from pluripotent stem cells (PSC). Materials and Methods Human PSCs were induced into SSCs. FACS and qRT-PCR were used to determine differences in expression of cell surface biomarkers and transcription factors between SSCs derived from PSCs and isolated from BM, in differentiating cells, in cells from early and late passage, and in fibroblasts. Results A significant reduction in proliferation and capacity of SSCs to differentiate into adipocytes and osteoblasts was observed after 3 passages. Protein and mRNA analysis indicated that commonly used biomarkers remain highly expressed in cells that lost capacity for differentiation. However, integrin alpha 6 (CD49f) and transcription factors GATA6, PRDM16, SIM2 and SOX11 were significantly upregulated in SSCs compared to fibroblasts. In early stages of adipogenic and osteogenic differentiation, the expression of CD49f, GATA6 and SIM2 was reduced in later passage cells, which have limited proliferation and differentiation capabilities. Conclusions Our results suggest that CD49f and transcription factors GATA6 and SIM2 identify functional SSCs.