Heo Yun Seok, Nagrath Sunitha, Moore Alessandra L, Zeinali Mahnaz, Irimia Daniel, Stott Shannon L, Toth Thomas L, Toner Mehmet
BioMEMS Resource Center, Mass achusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, MA.
BioMEMS Resource Center, Mass achusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, MA ; Massachusetts General Hospital, Cancer Center, Harvard Medical School, Boston, MA.
Technology (Singap World Sci). 2015 Mar;3(1):64-71. doi: 10.1142/S2339547815500053.
Long-term preservation of live cells is critical for a broad range of clinical and research applications. With the increasing diversity of cells that need to be preserved (e.g. oocytes, stem and other primary cells, genetically modified cells), careful optimization of preservation protocols becomes tedious and poses significant limitations for all but the most expert users. To address the challenge of long-term storage of critical, heterogeneous cell types, we propose a universal protocol for cell vitrification that is independent of cell phenotype and uses only low concentrations of cryoprotectant (1.5 M PROH and 0.5 M trehalose). We employed industrial grade microcapillaries made of highly conductive fused silica, which are commonly used for analytical chemistry applications. The minimal mass and thermal inertia of the microcapillaries enabled us to achieve ultrafast cooling rates up to 4,000 K/s. Using the same low, non-toxic concentration of cryoprotectant, we demonstrate high recovery and viability rates after vitrification for human mammary epithelial cells, rat hepatocytes, tumor cells from pleural effusions, and multiple cancer cell lines.
活细胞的长期保存对于广泛的临床和研究应用至关重要。随着需要保存的细胞种类日益多样（如卵母细胞、干细胞和其他原代细胞、基因改造细胞），仔细优化保存方案变得繁琐，而且对除最专业的用户之外的所有人都构成了重大限制。为应对关键的异质细胞类型长期储存的挑战，我们提出了一种通用的细胞玻璃化方案，该方案与细胞表型无关，且仅使用低浓度的冷冻保护剂（1.5 M 丙二醇和 0.5 M 海藻糖）。我们采用了由高导电性熔融石英制成的工业级微毛细管，这种微毛细管常用于分析化学应用。微毛细管的最小质量和热惯性使我们能够实现高达 4000 K/s 的超快冷却速率。使用相同的低浓度无毒冷冻保护剂，我们证明了人乳腺上皮细胞、大鼠肝细胞、胸腔积液肿瘤细胞和多种癌细胞系在玻璃化后的高回收率和活力率。
