Isolation of SSEA-3-positive mesenchymal stem cells from equine bone marrow and evaluation of their pluripotency

IF 2.2 3区农林科学 Q1 VETERINARY SCIENCES

Research in veterinary science Pub Date : 2025-06-03 DOI:10.1016/j.rvsc.2025.105736

Sangchan Lee , Mya Theingi Kyaw , Koji Harada , Ken Takeshi Kusakabe , Masaya Igase , Naoki Sasaki

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Abstract

Generally, most mesenchymal stem cells (MSCs) have lower pluripotency and limited differentiation potential than embryonic stem cells (ESCs). However, a small subpopulation of MSCs, called multilineage differentiating stress-enduring (MUSE) cells, exhibit pluripotency. MUSE cells express stage-specific embryonic antigen 3 (SSEA-3), a sphingoglycolipid. Here, we isolated and investigated the pluripotency of SSEA-3-positive MSCs (MUSE cells).

Six thoroughbred horses were used as test subjects. MSCs were harvested from the bone marrow of the thoracic vertebrae under ultrasound guidance. Harvested MSCs were sorted into SSEA-3-positive and -negative cells using a cell sorter, and their pluripotency was evaluated by immunostaining for the markers NANOG, OCT4, and SOX2.

The results showed that 2.1 % of equine MSCs were SSEA-3-positive, and these cells were successfully isolated using a cell sorter. This study confirms that SSEA-3-positive cells are present in equine bone marrow-derived MSCs, similar to that in other species.

Immunostaining revealed SOX2 (88.3 ± 11.6 %) and NANOG expressions (9.0 ± 3.9 %) in the SSEA-3-negative cell group, whereas OCT4 expression was not detected (0.0 ± 0.0 %). Contrastingly, the SSEA-3-positive cell group showed significant NANOG (19.8 ± 3.8 %), OCT4 (9.7 ± 2.6 %), and SOX2 expressions (97.3 ± 4.1 %), with significantly higher values for NANOG and OCT4 compared to the SSEA-3-negative group (p < 0.05).

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马骨髓ssea -3阳性间充质干细胞的分离及多能性评价

一般来说，大多数间充质干细胞（MSCs）的多能性和分化潜力都低于胚胎干细胞（ESCs）。然而，MSCs的一个小亚群，称为多谱系分化耐压（MUSE）细胞，表现出多能性。MUSE细胞表达阶段特异性胚胎抗原3 (SSEA-3)，一种鞘糖脂。在这里，我们分离并研究了ssea -3阳性MSCs （MUSE细胞）的多能性。6匹纯种马被用作试验对象。在超声引导下，从胸椎骨髓中获取MSCs。利用细胞分选器将收获的MSCs分为ssea -3阳性细胞和ssea -3阴性细胞，并通过NANOG、OCT4和SOX2标记物的免疫染色评估其多能性。结果显示，2.1%的马间充质干细胞为ssea -3阳性，这些细胞通过细胞分选器成功分离。本研究证实，ssea -3阳性细胞存在于马骨髓源性间充质干细胞中，与其他物种相似。免疫染色显示ssea -3阴性细胞组SOX2（88.3±11.6%）和NANOG（9.0±3.9%）表达，OCT4未见表达（0.0±0.0%）。相比之下，ssea -3阳性细胞组NANOG（19.8±3.8%）、OCT4（9.7±2.6%）和SOX2（97.3±4.1%）表达显著，且NANOG和OCT4表达显著高于ssea -3阴性细胞组(p <；0.05)。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Research in veterinary science 农林科学-兽医学

CiteScore

4.40

自引率

4.20%

发文量

312

审稿时长

75 days

期刊介绍： Research in Veterinary Science is an International multi-disciplinary journal publishing original articles, reviews and short communications of a high scientific and ethical standard in all aspects of veterinary and biomedical research. The primary aim of the journal is to inform veterinary and biomedical scientists of significant advances in veterinary and related research through prompt publication and dissemination. Secondly, the journal aims to provide a general multi-disciplinary forum for discussion and debate of news and issues concerning veterinary science. Thirdly, to promote the dissemination of knowledge to a broader range of professions, globally. High quality papers on all species of animals are considered, particularly those considered to be of high scientific importance and originality, and with interdisciplinary interest. The journal encourages papers providing results that have clear implications for understanding disease pathogenesis and for the development of control measures or treatments, as well as those dealing with a comparative biomedical approach, which represents a substantial improvement to animal and human health. Studies without a robust scientific hypothesis or that are preliminary, or of weak originality, as well as negative results, are not appropriate for the journal. Furthermore, observational approaches, case studies or field reports lacking an advancement in general knowledge do not fall within the scope of the journal.