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首页> 外文期刊>Journal of cellular biochemistry. >DMSO- and Serum-Free Cryopreservation of Wharton's Jelly Tissue Isolated From Human Umbilical Cord
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DMSO- and Serum-Free Cryopreservation of Wharton's Jelly Tissue Isolated From Human Umbilical Cord

机译：从人脐带分离的沃顿氏胶冻组织的DMSO和无血清冷冻保存

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摘要

The facile nature of mesenchymal stem cell (MSC) acquisition in relatively large numbers has made Wharton's jelly (WJ) tissue an alternative source of MSCs for regenerative medicine. However, freezing of such tissue using dimethyl sulfoxide (DMSO) for future use impedes its clinical utility. In this study, we compared the effect of two different cryoprotectants (DMSO and cocktail solution) on post-thaw cell behavior upon freezing of WJ tissue following two different freezing protocols (Conventional [-1 degrees C/min] and programmed). The programmed method showed higher cell survival rate compared to conventional method of freezing. Further, cocktail solution showed better cryoprotection than DMSO. Post-thaw growth characteristics and stem cell behavior of Wharton's jelly mesenchymal stem cells (WJMSCs) from WJ tissue cryopreserved with a cocktail solution in conjunction with programmed method (Prog-Cock) were comparable with WJMSCs from fresh WJ tissue. They preserved their expression of surface markers, pluripotent factors, and successfully differentiated in vitro into osteocytes, adipocytes, chondrocytes, and hepatocytes. They also produced lesser annexin-V-positive cells compared to cells from WJ tissue stored using cocktail solution in conjunction with the conventional method (Conv-Cock). Real-time PCR and Western blot analysis of post-thaw WJMSCs from Conv-Cock group showed significantly increased expression of pro-apoptotic factors (BAX, p53, and p21) and reduced expression of anti-apoptotic factor (BCL2) compared to WJMSCs from the fresh and Prog-Cock group. Therefore, we conclude that freezing of fresh WJ tissue using cocktail solution in conjunction with programmed freezing method allows for an efficient WJ tissue banking for future MSC-based regenerative therapies. J. Cell. Biochem. 117: 2397-2412, 2016. (c) 2016 Wiley Periodicals, Inc.
机译：间充质干细胞（MSC）的获取相对容易，使沃顿氏胶冻（WJ）组织成为再生医学MSC的替代来源。但是，使用二甲亚砜（DMSO）冷冻此类组织以备将来使用会妨碍其临床应用。在这项研究中，我们比较了两种不同的冷冻方案（常规[-1℃/ min]和程序设置）对WJ组织冷冻后两种不同的防冻剂（DMSO和鸡尾酒溶液）对融化后细胞行为的影响。与常规冷冻方法相比，编程方法显示出更高的细胞存活率。此外，鸡尾酒溶液显示出比DMSO更好的冷冻保护。结合了鸡尾酒溶液和程序化方法（Prog-Cock）冷冻保存的WJ组织的沃顿氏果冻间充质干细胞（WJMSC）的融化后生长特性和干细胞行为与新鲜WJ组织的WJMSC相当。他们保留了表面标志物，多能因子的表达，并在体外成功分化为骨细胞，脂肪细胞，软骨细胞和肝细胞。与使用鸡尾酒溶液结合常规方法（Conv-Cock）储存的WJ组织细胞相比，它们还产生了较少的膜联蛋白V阳性细胞。 Conv-Cock组解冻后WJMSC的实时PCR和Western印迹分析表明，与来自WJMSC的WJMSC相比，促凋亡因子（BAX，p53和p21）的表达显着增加，而抗凋亡因子（BCL2）的表达降低。新鲜和Prog-Cock组。因此，我们得出结论，使用鸡尾酒溶液结合程序冷冻方法冷冻新鲜的WJ组织，可以为未来基于MSC的再生疗法提供有效的WJ组织储备。 J.细胞。生化。 117：2397-2412，2016。（c）2016 Wiley Periodicals，Inc.

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