Amphipathic polymer-mediated uptake of trehalose for dimethyl sulfoxide-free human cell cryopreservation.
Amphipathic polymer-mediated uptake of trehalose for dimethyl sulfoxide-free human cell cryopreservation.
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DOI：
10.1016/j.cryobiol.2013.09.002
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发表时间：
2013-12
期刊：
影响因子：
2.7
通讯作者：
Slater, Nigel K. H.
中科院分区：
文献类型：
--
作者：
Sharp, Duncan M. C.;Picken, Andrew;Morris, Timothy J.;Hewitt, Christopher J.;Coopman, Karen;Slater, Nigel K. H.
为了使干细胞治疗成为常规现实，需要克服的主要挑战之一是它们的储存和运输。目前，这是通过利用冷冻保护剂二甲基亚砜（Me 2SO）冷冻保存细胞来实现的。Me 2SO对细胞有毒，导致细胞功能丧失，并可能对患者产生严重的副作用。如果能够以足够的浓度将冷冻保护剂海藻糖递送到细胞中，则可能使用冷冻保护剂海藻糖冷冻细胞。新的两亲性膜透化剂PP-50先前已被证明可增强红细胞对海藻糖的摄取，从而增加低温存活。在这里，这项工作扩展到有核人类细胞系SAOS-2。使用最佳PP-50浓度和培养基渗透压，解冻后细胞活力为60 ± 2%。此外，解冻后24小时代谢活性细胞的数量（标准化至冷冻前）在103 ± 4%和91 ± 5%之间。发现这与使用Me 2SO冷冻的细胞相当。尽管倍增时间缩短（22 ± 2%，p = 0.09），但与非冷冻对照相比无统计学差异。这与使用Me 2SO冷冻的细胞形成对比，其中倍增时间显著减少（41 ± 4%，p = 0.004）。PP-50介导的海藻糖递送到细胞中可以代表适合于研究和治疗应用的替代冷冻保存方案。
For stem cell therapy to become a routine reality, one of the major challenges to overcome is their storage and transportation. Currently this is achieved by cryopreserving cells utilising the cryoprotectant dimethyl sulfoxide (Me2SO). Me2SO is toxic to cells, leads to loss of cell functionality, and can produce severe side effects in patients. Potentially, cells could be frozen using the cryoprotectant trehalose if it could be delivered into the cells at a sufficient concentration. The novel amphipathic membrane permeabilising agent PP-50 has previously been shown to enhance trehalose uptake by erythrocytes, resulting in increased cryosurvival. Here, this work was extended to the nucleated human cell line SAOS-2. Using the optimum PP-50 concentration and media osmolarity, cell viability post-thaw was 60 ± 2%. In addition, the number of metabolically active cells 24 h post-thaw, normalised to that before freezing, was found to be between 103 ± 4% and 91 ± 5%. This was found to be comparable to cells frozen using Me2SO. Although reduced (by 22 ± 2%, p = 0.09), the doubling time was found not to be statistically different to the non-frozen control. This was in contrast to cells frozen using Me2SO, where the doubling time was significantly reduced (by 41 ± 4%, p = 0.004). PP-50 mediated trehalose delivery into cells could represent an alternative cryopreservation protocol, suitable for research and therapeutic applications.
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影响因子：
3.5
作者：
Adinolfi, Elena;Pizzirani, Cinzia;Idzko, Marco;Panther, Elisabeth;Norgauer, Johannes;Di Virgilio, Francesco;Ferrari, Davide
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Ferrari, Davide
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影响因子：
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作者：
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通讯作者：
Slater, Nigel K. H.
影响因子：
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