Long-Term Trypsin Treatment Promotes Stem Cell Potency of Canine Adipose-Derived Mesenchymal Stem Cells
Long-Term Trypsin Treatment Promotes Stem Cell Potency of Canine Adipose-Derived Mesenchymal Stem Cells
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DOI：
10.1089/scd.2020.0175
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发表时间：
2021-03-04
影响因子：
4
通讯作者：
Inaba，Toshio
中科院分区：
文献类型：
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作者：
Mitani，Kosuke;Ito，Yuki;Inaba，Toshio
从脂肪组织中分离的间充质干细胞(ADSCs)被认为是在再生医学中应用最有前途的细胞类型之一。然而，由于异质性，ADSCs的再生能力可能会有所不同。已知长期胰酶治疗(LTT)可显著浓缩人MSCs的多系分化应激耐受(MUSE)细胞。在这项研究中，我们的目标是利用LTT从犬ADSCs中培养出具有高干细胞潜能的细胞。经胰酶处理16 h后，存活的ADSCs(长期耐受细胞)的胚胎干细胞标志物阶段特异性胚胎抗原1和岩藻糖基转移酶9的表达显著增强。然而，LTT耐受细胞并没有增强SSEA-3的表达，SSEA-3是一种已知的人类缪斯细胞标志。然而，LTT耐受细胞在集落形成单位成纤维细胞实验中显示出显著高于ADSCs的自我更新能力。此外，耐LTT的细胞形成类似类胚体的细胞团，并表达未分化的标记。此外，这些细胞分化为所有三个生殖层的细胞，并显示出明显高于ADSC的α2-6唾液酸(SIA)特异性凝集素水平，这些凝集素是人MSCs的分化潜能标记物。LTT耐受细胞核型正常，端粒酶活性低，致癌能力较弱。与ADSCs相比，LTT耐受细胞在化学诱导剂存在下的迁移活性显著增强，迁移相关基因的表达增加。此外，LTT耐受细胞对丝裂原刺激的淋巴细胞增殖的抑制作用强于ADSCs。总体而言，这些结果表明，犬ADSCs中的LTT耐受细胞具有与人类MUSE细胞相似的特性(尽管其中一个未分化的标记不同)，有望成为狗的再生治疗的一种有前途的工具。
Mesenchymal stem cells (MSCs) isolated from adipose tissue (adipose-derived stem cells [ADSCs]) are considered one of the most promising cell types for applications in regenerative medicine. However, the regenerative potency of ADSCs may vary because of heterogeneity. Long-term trypsin treatment (LTT) is known to significantly concentrate multilineage-differentiating stress-enduring (Muse) cells from human MSCs. In this study, we aimed to generate cells with high stem cell potency from canine ADSCs using LTT. After 16 h of treatment with trypsin, surviving ADSCs (LTT-tolerant cells) had significantly enhanced expression of stage-specific embryonic antigen (SSEA)-1, a mouse embryonic stem cell marker, and fucosyltransferase 9, one of several fucosyltransferases for SSEA-1 biosynthesis. However, LTT-tolerant cells did not enhance the expression of SSEA-3, a known human Muse cell marker. LTT-tolerant cells, however, showed significantly higher self-renewal capacity in the colony-forming unit fibroblast assay than ADSCs. In addition, the LTT-tolerant cells formed cell clusters similar to embryoid bodies and expressed undifferentiated markers. Moreover, these cells differentiated into cells of all three germ layers and showed significantly higher levels of α 2–6 sialic acid (Sia)-specific lectins, known as differentiation potential markers of human MSCs, than ADSCs. LTT-tolerant cells had a normal karyotype and had low telomerase activity, showing little carcinogenetic potency. LTT-tolerant cells also showed significantly increased activity of transmigration in the presence of chemoattractants and had increased expression of migration-related genes compared with ADSCs. In addition, LTT-tolerant cells had stronger suppressive activity against mitogen-stimulated lymphocyte proliferation than ADSCs. Overall, these results indicated that the LTT-tolerant cells in canine ADSCs have similar properties as human Muse cells (although one of the undifferentiated markers is different) and are expected to be a promising tool for regenerative therapy in dogs.