Li Xin, Cao Yukun, Liu Chenxi, Tan Jia, Zhou Xinli
Institute of Biothermal Science & Technology, University of Shanghai for Science and Technology, Shanghai, 200093, China; Shanghai Co-innovation Center for Energy Therapy of Tumors, Shanghai, 200093, China; Shanghai Technical Service Platform for Cryopreservation of Biological Resources, Shanghai, 200093, China.
Institute of Biothermal Science & Technology, University of Shanghai for Science and Technology, Shanghai, 200093, China; Shanghai Co-innovation Center for Energy Therapy of Tumors, Shanghai, 200093, China; Shanghai Technical Service Platform for Cryopreservation of Biological Resources, Shanghai, 200093, China.
Cryobiology. 2024 Sep;116:104942. doi: 10.1016/j.cryobiol.2024.104942. Epub 2024 Jul 30.
Cryopreservation of biological samples is an important technology for expanding their applications in the biomedical field. However, the quality and functionality of samples after rewarming are limited by the toxicity of commonly used cryoprotectant agents (CPAs). Here, we developed a novel preservation system by combining the natural amino acid l-proline (L-Pro) with gelatin methacryloyl (GelMA) hydrogels. Compared with dimethyl sulfoxide (DMSO), L-Pro and GelMA demonstrated excellent biocompatibility when co-culturing with cells. Cryopreservation procedures were optimized using 3T3 as model cells. The results showed that rapid cooling was the most suitable cooling procedure for L-Pro and GelMA among the three cooling procedures. Co-culturing with cells for 3 h before cryopreservation, 6 % L-Pro +7 % GelMA had the highest survival rate, reaching up to 80 %. Differential Scanning Calorimetry (DSC) analysis showed that 6 % L-Pro + 7 % GelMA lowered the freezing point of the solution to -4.2 °C and increased the unfrozen water content to 20 %. To the best of our knowledge, this is the first report of cell cryopreservation using a combination of L-Pro and GelMA hydrogels, which provides a new strategy for improving cell cryopreservation.
生物样品的冷冻保存是扩大其在生物医学领域应用的一项重要技术。然而，复温后样品的质量和功能受到常用冷冻保护剂（CPA）毒性的限制。在此，我们通过将天然氨基酸L-脯氨酸（L-Pro）与甲基丙烯酰化明胶（GelMA）水凝胶相结合，开发了一种新型保存系统。与二甲基亚砜（DMSO）相比，L-Pro和GelMA在与细胞共培养时表现出优异的生物相容性。以3T3作为模型细胞对冷冻保存程序进行了优化。结果表明，在三种冷却程序中，快速冷却是最适合L-Pro和GelMA的冷却程序。在冷冻保存前与细胞共培养3小时，6%L-Pro + 7%GelMA具有最高的存活率，可达80%。差示扫描量热法（DSC）分析表明，6%L-Pro + 7%GelMA将溶液的冰点降低至-4.2°C，并将未冻水含量提高至20%。据我们所知，这是首次关于使用L-Pro和GelMA水凝胶组合进行细胞冷冻保存的报道，为改善细胞冷冻保存提供了一种新策略。
