Interfacial Interactions of Sucrose during Cryopreservation Detected by Raman Spectroscopy.
Interfacial Interactions of Sucrose during Cryopreservation Detected by Raman Spectroscopy.
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DOI：
10.1021/acs.langmuir.8b01616
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发表时间：
2019-06-11
期刊：
影响因子：
--
通讯作者：
Hubel A
中科院分区：
文献类型：
--
作者：
Yu G;Li R;Hubel A
人们对利用糖来保存细胞很感兴趣。在本研究中，低温拉曼光谱用于表征蔗糖在冷冻过程中的行为。在- 10°C和- 50°C时研究了蔗糖和水之间的氢键网络，拉曼光谱显示，在- 50°C时，更浓的蔗糖溶液中蔗糖-水和蔗糖-蔗糖氢键增强。在冰界面处，蔗糖浓度随着冰密度的降低而增加，并在一个狭窄的非冷冻蔗糖溶液通道上趋于稳定，然后向下一个冰界面下降。研究了细胞与非冷冻蔗糖溶液界面以及细胞与细胞外冰界面的生物物理环境。在细胞与细胞外冰的界面处观察到一层薄薄的非冷冻蔗糖溶液。该界面处的细胞外蔗糖浓度普遍低于散装非冷冻蔗糖溶液。细胞膜不同区域外蔗糖浓度的变化表明，在冷冻过程中，细胞周围的化学环境可能比以前认为的更不均匀。拉曼光谱和图像还显示了非冷冻蔗糖溶液与细胞的共定位，这表明蔗糖与细胞膜的直接相互作用可能是蔗糖具有保护作用的原因。蔗糖主要分布在细胞外，观察到蔗糖在细胞膜上的强烈分配与拉曼光谱检测到的大量细胞脱水一致。这项工作提高了我们对蔗糖溶液的行为及其与细胞在低温下的相互作用的理解，并可以改进细胞在蔗糖基培养基中冷冻的低温保存方案。
There is considerable interest in the use of sugars to preserve cells. In this study, low temperature Raman spectroscopy was used to characterize the behaviors of sucrose during freezing. The hydrogen bond network between sucrose and water was investigated at −10 °C and −50 °C, and the Raman spectra showed strengthened sucrose-water and sucrose-sucrose hydrogen bonds in more concentrated sucrose solution at −50 °C. The concentration of sucrose at the ice interface increased as the ice density decreased, and it plateaued across a narrow channel of nonfrozen sucrose solution before it decreased toward the next ice interface. The biophysical environment at interfaces between the cell and nonfrozen sucrose solution and between the cell and extracellular ice was also studied. A thin layer of nonfrozen sucrose solution was observed at the interface between the cell and extracellular ice. The extracellular concentration of sucrose at this interface was generally lower than that of bulk nonfrozen sucrose solution. The variation of sucrose concentration outside different regions of the cell membrane suggests that the chemical environment around the cell during freezing may be more heterogeneous than previously thought. Raman spectra and images also showed colocalization of nonfrozen sucrose solution and the cell, which implied that direct interaction between sucrose and cell membrane might be responsible for protective properties of sucrose. Sucrose was predominantly distributed outside the cell, and the observation of strong partitioning of sucrose across the cell membrane is consistent with substantial cell dehydration detected by the Raman spectra. This work enhances our understanding of the behaviors of sucrose solution and its interactions with cells at low temperature and can improve cryopreservation protocols of cells frozen in a sucrose-based media.
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影响因子：
3.3
作者：
Dashnau, Jennifer L.;Nucci, Nathaniel V.;Vanderkooi, Jane M.
通讯作者：
Vanderkooi, Jane M.
影响因子：
4.1
作者：
Chang, Anthony;Dantzig, Jonathan A.;Hubel, Allison
通讯作者：
Hubel, Allison
DOI：
10.1111/j.1749-6632.1998.tb10162.x
发表时间：
1998-01-01
期刊：
BIOTRANSPORT: HEAT AND MASS TRANSFER IN LIVING SYSTEMS
影响因子：
--
作者：
Harmison, HR;Diller, KR;Brand, JJ
通讯作者：
Brand, JJ
影响因子：
3.1
作者：
Beatriz Brizuela, Alicia;Cecilia Bichara, Laura;Antonia Brandan, Silvia
通讯作者：
Antonia Brandan, Silvia
影响因子：
3.4
作者：
Jaeger, M;Carin, M;Tryggvason, G
通讯作者：
Tryggvason, G