Generation of UCiPSC-derived neurospheres for cell therapy and its application

Stem Cell Research & Therapy

Huifang Zhao¹  Chao Tian¹  Feng Tang²  Zuoxian Lin³  Rongqi Huang³  Xiaobo Han³  Omar Mukama³  Shuai Li³  Hualin Huang³  Zhiyuan Li⁴  Lang He⁵  Bin Ni⁶  Jean de Dieu Habimana⁷ 

[1] CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China;Guangzhou Regenerative Medicine and Health Guangdong Laboratory, 510005, Guangzhou, China;GZMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou, China;CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China;Guangzhou Regenerative Medicine and Health Guangdong Laboratory, 510005, Guangzhou, China;GZMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou, China;Department of Anatomy and Neurobiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China;CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China;Guangzhou Regenerative Medicine and Health Guangdong Laboratory, 510005, Guangzhou, China;GZMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou, China;University of Chinese Academy of Sciences, 100049, Beijing, China;CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China;Guangzhou Regenerative Medicine and Health Guangdong Laboratory, 510005, Guangzhou, China;GZMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou, China;University of Chinese Academy of Sciences, 100049, Beijing, China;NHC Key Laboratory of Birth Defect for Research and Prevention, Hunan Provincial Maternal and Child Health Care Hospital, 410008, Changsha, Hunan, China;Department of Anatomy and Neurobiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China;Department of Anatomy and Neurobiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China;NHC Key Laboratory of Birth Defect for Research and Prevention, Hunan Provincial Maternal and Child Health Care Hospital, 410008, Changsha, Hunan, China;University of Chinese Academy of Sciences, 100049, Beijing, China;

关键词: Human induced pluripotent stem cells;    hiPSC-derived human neural stem cells;    Neurospheres;    Transportation;    Ambient temperature;   

DOI  :  10.1186/s13287-021-02238-4

来源: Springer

【 摘 要 】

BackgroundNeural stem cell (NSC) therapy remains one of the most potential approaches for the treatment of neurological disorders. The discovery of human induced pluripotent stem cells (hiPSCs) and the establishment of hiPSC-derived human neural stem cells (hiNSCs) have revolutionized the technique of cell therapy. Meanwhile, it is often required that NSCs are stored and transported to a long distance for research or treatment purposes. Although high survival rates could be maintained, conventional methods for cell transportation (dry ice or liquid nitrogen) are inconvenient and expensive. Therefore, the establishment of a safe, affordable, and low-cost strategy to store and transport easily accessible hiPSCs and hiNSCs, with characteristics that match fetal hNSCs, is incredibly urgent.MethodsWe reprogrammed human urinary cells to iPSCs using a non-integrating, virus-free technique and differentiated the iPSCs toward iNSCs/neurospheres and neurons, under Good Manufacturing Practice (GMP)-compatible conditions. The pluripotency of iPSCs and iNSCs was characterized by a series of classical methods (surface markers, karyotype analysis, and in vitro as well as in vivo differentiation capabilities, etc.).ResultsHere, our results showed that we successfully generated hiNSCs/neurospheres from more available, non-invasive, and more acceptable urinary cells by a virus-free technique. Next, we demonstrated that the iNSCs differentiated into mature cerebral cortical neurons and neural networks. Interestingly, hiNSCs survived longer as neurospheres at ambient temperature (AT) than those cultured in a monolayer. Within 7 days approximately, the neural viability remained at > 80%, while hiNSCs cultured in a monolayer died almost immediately. Neurospheres exposed to AT that were placed under standard culture conditions (37 °C, 5% CO2) recovered their typical morphology, and retained their proliferation and differentiation abilities.ConclusionsIn this study, we provided a simple method for the storage of NSCs as neurospheres at AT as an alternative method to more costly and inconvenient traditional methods of cryopreservation. This will enable hiNSCs to be transported over long distances at AT and facilitate the therapeutic application of NSCs as neurospheres without any further treatment.

【 授权许可】

CC BY