Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, First Affiliated Hospital of Hainan Medical University, Hainan Medical University, Haikou, China; Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou, China; Hainan Provincial Clinical Research Center for Thalassemia, First Affiliated Hospital of Hainan Medical University, Hainan Medical University, Haikou, China.
Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, First Affiliated Hospital of Hainan Medical University, Hainan Medical University, Haikou, China.
Cryobiology. 2021 Feb;98:210-214. doi: 10.1016/j.cryobiol.2020.10.019. Epub 2020 Nov 2.
Human induced pluripotent stem cells (hiPSCs) have the properties of differentiation potential and unlimited self-renewal. Developing efficient and highly safe methods to preserve hiPSCs is important due to they have demonstrated tremendous promise in disease etiology, drug discovery, and regenerative medicine applications. Traditionally, open systems for cell cryopreservation, such as conventional slow freezing and vitrification methods, were widespread application in the storage and transportation of hiPSCs. However, these two methods have such problems of low recovery rate and the risk of cross-contamination. Recently, closed systems for cell cryopreservation, such as CryoLogic Vitrification Method (CVM), were introduced to store and transport embryos. In this study, we developed a new friendly CVM by loading a small piece of hiPSCs colonies in the vitrification solution to the hook of Fiberplug to increase the cooling rate. To warm them, the CVM Fiberplug was immersed directly in a 37 °C warming solution for 1 min, and hiPSCs were then transferred to mTeSR1 medium. The result revealed that the new CVM had a high recovery rate and maintained the stemness and differentiation potential of hiPSCs. Our new CVM not only provide a safe way for hiPSCs preservation but also has a high survival rate in the storage of hiPSCs.
人诱导多能干细胞（hiPSCs）具有分化潜能和无限自我更新的特性。由于 hiPSCs 在疾病发病机制、药物发现和再生医学应用方面表现出巨大的潜力，因此开发高效、高度安全的方法来保存 hiPSCs 非常重要。传统上，细胞冷冻保存的开放式系统，如传统的慢速冷冻和玻璃化方法，广泛应用于 hiPSCs 的储存和运输。然而，这两种方法存在回收率低和交叉污染风险等问题。最近，细胞冷冻保存的封闭式系统，如 CryoLogic Vitrification Method（CVM），已被引入用于储存和运输胚胎。在本研究中，我们通过将一小片 hiPSCs 集落加载到 Fiberplug 的钩子上，然后将其放入玻璃化溶液中，开发了一种新的友好型 CVM，以提高冷却速度。为了对其进行加热，将 CVM Fiberplug 直接浸入 37°C 的温育溶液中 1 分钟，然后将 hiPSCs 转移到 mTeSR1 培养基中。结果表明，新型 CVM 具有较高的回收率，且维持了 hiPSCs 的干性和分化潜能。我们的新型 CVM 不仅为 hiPSCs 的保存提供了一种安全的方法，而且在 hiPSCs 的储存中也具有较高的存活率。
