在马脂肪组织间充质干细胞冷冻溶液中添加合成聚合物作为降低二甲基亚砜（DMSO）浓度的未来研究方向。
Addition of synthetic polymer in the freezing solution of mesenchymal stem cells from equine adipose tissue as a future perspective for reducing of DMSO concentration.
作者信息
Nascimento Cátia, Saraiva Márcia Viviane Alves, Pereira Vitoria Mattos, de Brito Danielle Cristina Calado, de Aguiar Francisco Léo Nascimento, Alves Benner Geraldo, Roballo Kelly Cristine Santos, de Figueiredo José Ricardo, Ambrósio Carlos Eduardo, Rodrigues Ana Paula Ribeiro
机构信息
Veterinarian, MSc. Laboratório de Manipulação de Oócitos e Folículos Pré-Antrais Ovarianos (LAMOFOPA), Faculdade de Medicina Veterinária, Universidade Estadual do Ceará, Fortaleza, CE, Brazil.
Veterinarian, DSc. Universidade Federal Rural do Semiárido (UFERSA), Mossoró, RN, Brazil.
出版信息
Braz J Vet Med. 2023 Dec 27;45:e002523. doi: 10.29374/2527-2179.bjvm002523. eCollection 2023.
The regenerative therapies with stem cells (SC) has been increased by the cryopreservation, permitting cell storage for extended periods. However, the permeating cryoprotectant agents (CPAs) such as dimethylsulfoxide (DMSO) can cause severe adverse effects. Therefore, this study evaluated equine mesenchymal stem cells derived from adipose tissue (eAT-MSCs) in fresh (Control) or after slow freezing (SF) in different freezing solutions (FS). The FS comprise DMSO and non-permeating CPAs [Trehalose (T) and the SuperCool X-1000 (X)] in association or not, totalizing seven different FS: (DMSO; T; X; DMSO+T; DMSO+X; T+X, and DMSO+T+X). Before and after cryopreservation were evaluated, viability, colony forming unit (CFU), and cellular differentiation capacity. After freezing-thawing, the viability of the eAT-MSCs reduced (P< 0.05) in all treatments compared to the control. However, the viability of frozen eAT-MSCs in DMSO (80.3 ± 0.6) was superior (P<0.05) to the other FS. Regarding CFU, no difference (P>0.05) was observed between fresh and frozen cells. After freezing-thawing, the eAT-MSCs showed osteogenic, chondrogenic, and adipogenic lineages differentiation potential. Nonetheless, despite the significative reduction in the osteogenic differentiation capacity between fresh and frozen cells, no differences (P > 0.05) were observed among FS. Furthermore, the number of chondrogenic differentiation cells frozen in DMSO+X solution reduced (P<0.05) comparing to the control, without differ (P>0.05) to the other FS. The adipogenic differentiation did not differ (P>0.05) among treatments. In conclusion, although these findings confirm the success of DMSO to cryopreserve eAT-MSCs, the Super Cool X-1000 could be a promise to reduce the DMSO concentration in a FS.
干细胞（SC）的再生疗法因冷冻保存技术而有所增加，这使得细胞能够长期储存。然而，诸如二甲亚砜（DMSO）等渗透性冷冻保护剂（CPA）会引发严重的不良反应。因此，本研究评估了源自脂肪组织的马间充质干细胞（eAT-MSCs），分别为新鲜状态（对照）以及在不同冷冻溶液（FS）中缓慢冷冻（SF）后的情况。FS包括DMSO和非渗透性CPA[海藻糖（T）和SuperCool X-1000（X）]，它们单独或联合使用，共有七种不同的FS：（DMSO；T；X；DMSO+T；DMSO+X；T+X，以及DMSO+T+X）。在冷冻保存前后，对细胞活力、集落形成单位（CFU）和细胞分化能力进行了评估。冻融后，与对照相比，所有处理组中eAT-MSCs的活力均降低（P<0.05）。然而，DMSO中冷冻的eAT-MSCs的活力（80.3±0.6）优于（P<0.05）其他FS。关于CFU，新鲜细胞和冷冻细胞之间未观察到差异（P>0.05）。冻融后，eAT-MSCs显示出成骨、软骨生成和脂肪生成谱系的分化潜能。尽管如此，尽管新鲜细胞和冷冻细胞之间的成骨分化能力有显著降低，但在FS之间未观察到差异（P>0.05）。此外，与对照相比，在DMSO+X溶液中冷冻的软骨生成分化细胞数量减少（P<0.05），与其他FS相比无差异（P>0.05）。脂肪生成分化在各处理组之间无差异（P>0.05）。总之，尽管这些发现证实了DMSO冷冻保存eAT-MSCs的成功，但SuperCool X-1000有望降低FS中DMSO的浓度。
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