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首页> 外文期刊>Cell biology international. >Effect of ectopic OCT4 expression on canine adipose tissue-derived mesenchymal stem cell proliferation
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Effect of ectopic OCT4 expression on canine adipose tissue-derived mesenchymal stem cell proliferation

机译：异位OCT4表达对犬脂肪组织衍生间充质干细胞增殖的影响

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摘要

Enhancing the proliferative capacity of mesenchymal stem cells (MSCs) is critical for increasing their therapeutic potential in a variety of diseases. We hypothesized that lentivirus-mediated overexpression of canine octamer-binding transcription factor 4 (OCT4) might influence the proliferation of canine adipose tissue-derived MSCs (cATMSCs). cOCT4-cATMSCs were generated by transducing cATMSCs with a cOCT4-lentiviral vector. Increased expression of cOCT4 was confirmed using RT-PCR and immunoblotting. Immunophenotypic characterization using flow cytometry indicated that the CD29, CD44, CD73, CD90, and CD 105 surface markers were highly expressed by both cOCT4- and mock-transduced cATMSCs (mock-cATMSCs), whereas the CD31 and CD45 markers were absent. We performed the osteogenic differentiation assay to evaluate the effects of cOCT4 overexpression on the osteogenic differentiation potential of cATMSCs. The results showed that cOCT4-cATMSCs had a much higher potential for osteogenic differentiation than mock-cATMSCs. Next, the proliferative capacities of cOCT4- and mock-cATMSCs were evaluated using a WST-1 cell proliferation assay and trypan blue exclusion. cOCT4-cATMSCs showed a higher proliferative capacity than mock-cATMSCs. Cell cycle analysis indicated that overexpression of cOCT4 in cATMSCs induced an increase in the proportion of cells in S and G2/M phases. Consistent with this, immunoblot analysis showed that cyclin D1 expression was increased in cOCT4-cATMSCs. In conclusion, our results indicate that lentivirus-mediated overexpression of cOCT4 increased the proliferative capacity of cATMSCs. OCT4-mediated enhancement of cell proliferation may be a useful method for expanding MSC population rapidly without loss of sternness.
机译：增强间充质干细胞（MSCs）的增殖能力对于增加各种疾病的治疗潜力至关重要。我们假设甘霉菌介导的犬八胞偶联转录因子4（OCT4）的过表达可能影响犬脂肪组织衍生的MSCs（Catmscs）的增殖。通过用COCT4-慢病毒载体转换CATMSS产生COCT4-CATMSC。使用RT-PCR和免疫印迹证实了COCT4的增加。使用流式细胞术的免疫蛋白酶表征表明CD29，CD44，CD73，CD90和CD 105表面标志物高度表达COCT4-和模型转导的CATMSCs（模拟-CTMSCs），而CD31和CD45标记不存在。我们进行了成骨分化测定以评估COCT4过表达对Catmscs的成骨分化电位的影响。结果表明，COCT4-Catmscs比模仿型锥形分化具有更高的骨质潜力。接下来，使用WST-1细胞增殖测定和台盼蓝排除来评估COCT4和模拟CATMSCs的增殖能力。 CoCT4-Catmscs显示出比模拟Catmscs更高的增殖能力。细胞循环分析表明CATMSCS中COCT4的过度表达诱导S和G2 / m相中细胞比例的增加。与此一致，免疫斑分析表明COCT4-Catmscs中的细胞周期蛋白D1表达增加。总之，我们的结果表明，慢病毒介导的COCT4的过表达增加了CATMSCs的增殖能力。 Oct4介导的细胞增殖的增强可能是扩展MSC种群的有用方法，而不会损失严厉。

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