| Institution: | 1. State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China;2. University of Chinese Academy of Sciences, Beijing 100049, PR China;1. School of Marine Science and Technology, Harbin Institute of Technology at Weihai, Weihai 264209, China;2. School of Municipal and Environmental Engineering, Harbin Institute of Technology, Harbin 150090, China;3. State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology, Harbin 150090, China;4. School of Environmental Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240, China;1. State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China;2. University of Chinese Academy of Sciences, Beijing 100049, PR China;3. Shuanglu Pharmaceutical Co., Ltd., No. 1 Building, Bitongyuan, No. 69 Fushi Road, Haidian District, Beijing 100043, PR China;1. Key Laboratory of Green Process and Engineering, National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China;2. University of Chinese Academy of Sciences, Beijing 100190, China;1. MOE Key Laboratory of Advanced Micro-structured Materials, School of Physics Science and Engineering, Tongji University, Shanghai 200092, China;2. School of Materials Science and Engineering, Jiangsu Key Laboratory for Advanced Metallic Materials, Southeast University, Nanjing 211189, China;3. Institute of Massive Amorphous Metal Science, China University of Mining and Technology, Xuzhou 221116, China |
| Abstract: | Low cell recovery rate of human embryonic stem cells (hESCs) resulting from cryopreservation damages leads to the difficulty in their successful commercialization of clinical applications. Hence in this study, sensitivity of human embryonic stem cells (hESCs) to different cooling rates, ice seeding and cryoprotective agent (CPA) types was compared and cell viability and recovery after cryopreservation under different cooling conditions were assessed. Both extracellular and intracellular ice formation were observed. Reactive oxidative species (ROS) accumulation of hESCs was determined. Cryopreservation of hESCs at 1 °C/min with the ice seeding and at the theoretically predicted optimal cooling rate (TPOCR) led to lower level of intracellular ROS, and prevented irregular and big ice clump formation compared with cryopreservation at 1 °C/min. This strategy further resulted in a significant increase in the hESC recovery when glycerol and 1,2-propanediol were used as the CPAs, but no increase for Me2SO. hESCs after cryopreservation under all the tested conditions still maintained their pluripotency. Our results provide guidance for improving the hESC cryopreservation recovery through the combination of CPA type, cooling rate and ice seeding. |
