Center for Engineering in Medicine and BioMEMS Resource Center, Massachusetts General Hospital, Harvard Medical School and Shriners Hospital for Children, Boston, Massachusetts, United States of America.
PLoS One. 2011;6(9):e24916. doi: 10.1371/journal.pone.0024916. Epub 2011 Sep 22.
This study reports an alternative approach to achieve vitrification where cells are pre-desiccated prior to cooling to cryogenic temperatures for storage. Chinese Hamster Ovary (CHO) cells suspended in a trehalose solution were rapidly and uniformly desiccated to a low moisture content (<0.12 g of water per g of dry weight) using a spin-drying technique. Trehalose was also introduced into the cells using a high-capacity trehalose transporter (TRET1). Fourier Transform Infrared Spectroscopy (FTIR) was used to examine the uniformity of water concentration distribution in the spin-dried samples. 62% of the cells were shown to survive spin-drying in the presence of trehalose following immediate rehydration. The spin-dried samples were stored in liquid nitrogen (LN(2)) at a vitrified state. It was shown that following re-warming to room temperature and re-hydration with a fully complemented cell culture medium, 51% of the spin-dried and vitrified cells survived and demonstrated normal growth characteristics. Spin-drying is a novel strategy that can be used to improve cryopreservation outcome by promoting rapid vitrification.
本研究报告了一种替代方法来实现玻璃化，即在将细胞冷却至低温储存之前对其进行预干燥。使用旋转干燥技术，将悬浮在海藻糖溶液中的中国仓鼠卵巢（CHO）细胞快速且均匀地干燥至低水分含量（<0.12 克水/克干重）。海藻糖也通过高容量海藻糖转运蛋白（TRET1）被引入细胞内。傅里叶变换红外光谱（FTIR）用于检查旋转干燥样品中水分浓度分布的均匀性。结果表明，在存在海藻糖的情况下，62%的细胞在立即复水后能够在旋转干燥中存活。旋转干燥的样品以玻璃化状态储存在液氮（LN(2)）中。结果表明，在室温下重新加热并使用完全补充的细胞培养基重新水合后，51%的旋转干燥和玻璃化细胞存活并表现出正常的生长特征。旋转干燥是一种新颖的策略，通过促进快速玻璃化可以提高冷冻保存的效果。
