Tateno Hiroaki, Saito Sayoko, Hiemori Keiko, Kiyoi Kayo, Hasehira Kayo, Toyoda Masashi, Onuma Yasuko, Ito Yuzuru, Akutsu Hidenori, Hirabayashi Jun
Biotechnology Research Institute for Drug Discovery (BRD), National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 2, 1-1-1 Umezono, Tsukuba, Ibaraki 305-8568, Japan
Biotechnology Research Institute for Drug Discovery (BRD), National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 2, 1-1-1 Umezono, Tsukuba, Ibaraki 305-8568, Japan.
Glycobiology. 2016 Dec;26(12):1328-1337. doi: 10.1093/glycob/cww039. Epub 2016 Apr 2.
Human somatic stem cells such as human mesenchymal stem cells (hMSCs) are considered attractive cell sources for stem cell-based therapy. However, quality control issues have been raised concerning their safety and efficacy. Here we used lectin microarray technology to identify cell surface glycans as markers of the differentiation potential of stem cells. We found that α2-6Sia-specific lectins show stronger binding to early passage adipose-derived hMSCs (with differentiation ability) than late passage cells (without the ability to differentiate). Flow cytometry analysis using α2-6Sia-specific lectins supported the results obtained by lectin microarray. Similar results were obtained for bone marrow-derived hMSCs and cartilage tissue-derived chondrocytes. Little or no binding of α2-6Sia-specific lectins was observed for human dermal fibroblasts, which are unable to differentiate, suggesting that the binding of α2-6Sia-specific lectins is associated with the differentiation ability of cells, but not to their capacity to proliferate. Quantitative analysis of the linkage mode of Sia using anion-exchange chromatography showed that the percentage of α2-6Sia linkage type was higher in early passage adipose-derived hMSCs than late passage cells. Integrinα5 was found to be a carrier protein of α2-6Sia. Sialidase treatment significantly reduced the differentiation efficiency of bone marrow-derived hMSCs. Based on these findings, we propose that α2-6sialylation is a marker of differentiation potential in stem cells such as adipose-derived hMSCs, bone marrow-derived hMSCs, and cartilage tissue-derived chondrocytes.
人类间充质干细胞（hMSCs）等人体体细胞干细胞被认为是基于干细胞治疗的有吸引力的细胞来源。然而，关于其安全性和有效性的质量控制问题已经被提出。在这里，我们使用凝集素微阵列技术来鉴定细胞表面聚糖作为干细胞分化潜能的标志物。我们发现，α2-6唾液酸特异性凝集素与早期传代的脂肪来源的hMSCs（具有分化能力）的结合比晚期传代细胞（无分化能力）更强。使用α2-6唾液酸特异性凝集素的流式细胞术分析支持了凝集素微阵列获得的结果。骨髓来源的hMSCs和软骨组织来源的软骨细胞也得到了类似的结果。对于无法分化的人皮肤成纤维细胞，未观察到α2-6唾液酸特异性凝集素的结合或仅有少量结合，这表明α2-6唾液酸特异性凝集素的结合与细胞的分化能力相关，而与其增殖能力无关。使用阴离子交换色谱对唾液酸连接方式的定量分析表明，早期传代的脂肪来源的hMSCs中α2-6唾液酸连接类型的百分比高于晚期传代细胞。整合素α5被发现是α2-6唾液酸的载体蛋白。唾液酸酶处理显著降低了骨髓来源的hMSCs的分化效率。基于这些发现，我们提出α2-6唾液酸化是脂肪来源的hMSCs、骨髓来源的hMSCs和软骨组织来源的软骨细胞等干细胞分化潜能的标志物。
