Cryoprotective enhancing effect of very low concentration of trehalose on the functions of primary rat hepatocytes.
Cryoprotective enhancing effect of very low concentration of trehalose on the functions of primary rat hepatocytes.
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DOI：
10.1016/j.reth.2020.08.003
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发表时间：
2020-12
影响因子：
4.3
通讯作者：
Ijima H
中科院分区：
文献类型：
--
作者：
Yoshida K;Ono F;Chouno T;Perocho BR;Ikegami Y;Shirakigawa N;Ijima H
细胞在生物医学研究中有各种应用。冷冻保存是一种细胞保存技术，可为此类应用提供细胞。冷冻保存后，敏感细胞，如原代肝细胞，由于冰晶引起的物理损伤而遭受低活力，这突出了对更好的冷冻保存方法以提高细胞活力的需要。考虑到有效抑制冰晶形成以保护细胞结构的重要性，海藻糖基于其抑制冰晶形成的能力作为冷冻保护剂引起了关注;然而，海藻糖诱导渗透胁迫。因此，要建立细胞冷冻保存技术，必须在海藻糖的保护作用和破坏作用之间提供最佳平衡。在这项研究中，我们评估了渗透胁迫和冰晶形成对原代大鼠肝细胞的活力和功能的影响，在较宽的海藻糖浓度范围内。在极低浓度（2.6 μM）的海藻糖下没有渗透胁迫，2.6 μM海藻糖驱动更细冰晶的形成，这对细胞膜的损伤较小。此外，我们发现在补充2.6 μM海藻糖的条件下冷冻保存后的活肝细胞数量比补充二甲基亚砜的条件下高70%。此外，未冻存的细胞和用海藻糖冻存的细胞显示出相当的细胞内脱氢酶活性。我们发现，在非常低的浓度（2.6 μM）的海藻糖显着提高冻存后的肝细胞的活力和肝功能。极低浓度的海藻糖可抑制冰晶形成，保护细胞结构。海藻糖的渗透压与肝细胞活力之间存在相关性。极低浓度的海藻糖可提高冻存后肝细胞的存活率和肝功能。
Cells have various applications in biomedical research. Cryopreservation is a cell-preservation technique that provides cells for such applications. After cryopreservation, sensitive cells, such as primary hepatocytes, suffer from low viability due to the physical damage caused by ice crystals, highlighting the need for better methods of cryopreservation to improve cell viability. Given the importance of effectively suppressing ice crystal formation to protect cellular structure, trehalose has attracted attention as cryoprotectant based on its ability to inhibit ice crystal formation; however, trehalose induces osmotic stress. Therefore, to establish a cell-cryopreservation technique, it is necessary to provide an optimal balance between the protective and damaging effects of trehalose. In this study, we evaluated the effects of osmotic stress and ice crystal formation on the viability and function of primary rat hepatocytes at wide range of trehalose concentration. There was no osmotic stress at very low concentrations (2.6 μM) of trehalose, and 2.6 μM trehalose drives the formation of finer ice crystals, which are less damaging to the cell membrane. Furthermore, we found that the number of viable hepatocytes after cryopreservation were 70% higher under the 2.6 μM trehalose-supplemented conditions than under the dimethyl sulfoxide-supplemented conditions. Moreover, non-cryopreserved cells and cells cryopreserved with trehalose showed comparable intracellular dehydrogenase activity. We showed that trehalose at very low concentrations (2.6 μM) improved dramatically viability and liver function of hepatocyte after cryopreservation. Very low concentration of trehalose could suppress ice crystal formation and protect cell structure. There was a correlation between osmotic pressure of trehalose and hepatocytes viability. Very low concentration of trehalose improved viability and liver function of hepatocyte after cryopreservation.
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影响因子：
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作者：
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通讯作者：
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