Next Generation Cell Cryopreservation: Developing Products for the Research Sector

创新细胞疗法和诊断、组织工程和干细胞库的市场正在迅速增长。细胞不能无限期地培养，因此必须冷冻保存以长期储存和分发。目前冷冻保存的黄金标准是使用高浓度的有毒有机溶剂，这是一项已有50年历史的技术，需要为21世纪的医药和工业进行更新。目前基于溶剂的方法不能完全恢复细胞，具有细胞毒性，而且许多细胞系目前根本不能冷冻储存。这导致了使用经过改造的耐寒细胞，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，而不是高质量但更脆弱的原代细胞。此外，新兴的细胞疗法（如CAR-T，一种革命性的癌症治疗方法，是英国第二大死亡原因）涉及将有毒的冷冻保存剂与细胞一起直接注射到患者体内。这会导致临床副作用，比如癌症患者变得非常虚弱，在某些情况下，他们的身体虚弱到必须暂停化疗或其他治疗的程度，使患者处于极度危险之中。为了应对这一挑战，我们从大自然中汲取了灵感。极端微生物（青蛙、蝎子和北极鱼）通过产生“抗冻蛋白”来生存，这种蛋白质可以减轻冰晶生长造成的损害，使它们能够在零度以下的环境中生存。CryoLogyx的研究团队开发了屡获殊荣（RSC-Emerging-Tech-2014）的合成大分子冷冻保护剂，这种聚合物通过模仿天然抗冻蛋白的某些功能，极大地改善了细胞的冷冻保存。该聚合物成本低，合成可扩展，并且已被证明可以显著提高从血液、细胞单层到干细胞等一系列细胞的解冻后细胞恢复。我们还将这些聚合物用于高价值抗体和酶的无溶剂储存。这是一种平台技术，在药物发现、高通量筛选、生物制剂和基础研究的所有领域都具有巨大潜力。它将支持英国生物技术，这是英国在世界上最好的产业之一，并将创造高技能的就业机会，并在中部地区和整个英国支持高科技制造业、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.