Next Generation Cell Cryopreservation: Developing Products for the Research Sector

创新的基于细胞的治疗和诊断、组织工程和干细胞库的市场正在迅速增长。细胞不能无限期培养，因此必须进行超低温保存，以便长期储存和分配。目前冷冻保存的黄金标准是使用高浓度有毒有机溶剂，这是一项有50年历史的技术，需要更新以适应21世纪的医学和工业。目前基于溶剂的方法不能完全回收细胞，具有细胞毒性，目前许多细胞系根本不能冷冻保存。这导致了工程化耐寒细胞的使用，这些细胞可以在解冻后重新生长。这些所谓的永生化细胞系因其健壮性而主导着研究领域，而不是更高质量但更脆弱的原代细胞。此外，新兴的基于细胞的疗法(如CAR-T，一种革命性的癌症治疗方法，在英国是第二大死亡原因)涉及将有毒的冷冻保护剂与细胞一起直接注入患者体内。这会导致临床副作用，比如癌症患者病情严重，在某些情况下，这些副作用会减弱到不得不暂停化疗或其他治疗的程度，将患者置于极端危险的境地。为了应对这一挑战，我们从自然中获得了灵感。极端微生物(从青蛙到蝎子再到北极鱼)通过产生“抗冻蛋白”来生存，这种蛋白可以减轻冰晶生长造成的损害，使它们能够生活在零度以下的环境中。我们在CryoLogyx的研究团队开发了获奖的(RSC-Emerging-Tech-2014)合成高分子低温保护剂--这种聚合物通过模仿天然抗冻蛋白的一些功能，显著改善了细胞的低温保存。这种聚合物成本低，可合成扩展，已被证明可显著提高从血液、细胞单层到干细胞等一系列细胞的解冻后细胞回收率。我们还将这些聚合物用于高价值抗体和酶的无溶剂存储。这是一种在药物发现、高通量筛选、生物制品和基础研究的所有领域都具有巨大潜力的平台技术。它将支持英国生物技术行业，这是英国的一个行业，是世界上最好的行业之一，并将创造高技能就业机会，并支持中部地区和整个英国的高科技制造业、运输和物流工作。

The market for innovative cell-based therapeutics and diagnostics, tissue engineering and stem cell banking is rapidly growing. Cells cannot be cultured indefinitely so must be cryopreserved for long term storage and distribution. The current gold standard for cryopreservation uses toxic organic solvents at high concentrations -- a 50 year old technology which needs updating for 21st century medicine and industry.The current solvent based approach does not give full cell recovery, is cytotoxic, and many cell lines simply cannot be stored frozen at present. This has led to the use of engineered hardy cells which can be regrown after thawing. These so-called immortalised cell lines dominate the research space due to their robustness, in preference to higher-quality, but more fragile, primary cells. Furthermore, emerging cell based therapies (such as CAR-T, a revolutionary treatment for cancer; the 2nd largest cause of death in the UK) involve the direct injection of toxic cryopreservatives into the patient alongside the cells. This leads to clinical side effects, such as the cancer patient becoming incredibly sick, and in some cases weakens them to the point where chemotherapy or other treatments have to be suspended, putting the patient in extreme danger.To address this challenge, we have taken inspiration from Nature. Extremophiles (frogs to scorpions to arctic fish) survive extreme cold by producing 'antifreeze proteins' which mitigate the damage caused by growing ice crystals, allowing them to live in sub-zero environments.Our research team at CryoLogyx have developed Award-winning (RSC-Emerging-Tech-2014) Synthetic Macromolecular Cryoprotectants -- polymers which dramatically improve cell cryopreservation by mimicking some functions of natural antifreeze proteins. The polymers are low cost, synthetically scalable and have been proven to dramatically increase post-thaw cell recovery for a range of cells, from blood, cell monolayers to stem cells. We have also used these polymers for the solvent-free storage of high-value antibodies and enzymes.This is a platform technology with huge potential across all areas of drug discovery, high throughput screening, biologics and fundamental research. It will support British Biotech, a UK industry that is one of the best in the world, and will create high-skilled jobs and support high tech manufacturing, transport and logistics jobs in the Midlands and across the UK.