Generation of UCiPSC-derived neurospheres for cell therapy and its application
Abstract Background Neural 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...
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| Format: | Article |
| Language: | English |
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BMC
2021-03-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13287-021-02238-4 |
| _version_ | 1818862605387694080 |
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| author | Shuai Li Huifang Zhao Xiaobo Han Bin Ni Lang He Omar Mukama Jean de Dieu Habimana Zuoxian Lin Rongqi Huang Hualin Huang Chao Tian Feng Tang Zhiyuan Li |
| author_facet | Shuai Li Huifang Zhao Xiaobo Han Bin Ni Lang He Omar Mukama Jean de Dieu Habimana Zuoxian Lin Rongqi Huang Hualin Huang Chao Tian Feng Tang Zhiyuan Li |
| author_sort | Shuai Li |
| collection | DOAJ |
| description | Abstract Background Neural 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. Methods We 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.). Results Here, 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. Conclusions In 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. |
| first_indexed | 2024-12-19T10:02:31Z |
| format | Article |
| id | doaj.art-c1e49df603cf47d0b4e3ff4ee31770f3 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-19T10:02:31Z |
| publishDate | 2021-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-c1e49df603cf47d0b4e3ff4ee31770f32022-12-21T20:26:36ZengBMCStem Cell Research & Therapy1757-65122021-03-0112111110.1186/s13287-021-02238-4Generation of UCiPSC-derived neurospheres for cell therapy and its applicationShuai Li0Huifang Zhao1Xiaobo Han2Bin Ni3Lang He4Omar Mukama5Jean de Dieu Habimana6Zuoxian Lin7Rongqi Huang8Hualin Huang9Chao Tian10Feng Tang11Zhiyuan Li12CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesNHC Key Laboratory of Birth Defect for Research and Prevention, Hunan Provincial Maternal and Child Health Care HospitalDepartment of Anatomy and Neurobiology, Xiangya School of Medicine, Central South UniversityCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesUniversity of Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesAbstract Background Neural 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. Methods We 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.). Results Here, 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. Conclusions In 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.https://doi.org/10.1186/s13287-021-02238-4Human induced pluripotent stem cellshiPSC-derived human neural stem cellsNeurospheresTransportationAmbient temperature |
| spellingShingle | Shuai Li Huifang Zhao Xiaobo Han Bin Ni Lang He Omar Mukama Jean de Dieu Habimana Zuoxian Lin Rongqi Huang Hualin Huang Chao Tian Feng Tang Zhiyuan Li Generation of UCiPSC-derived neurospheres for cell therapy and its application Stem Cell Research & Therapy Human induced pluripotent stem cells hiPSC-derived human neural stem cells Neurospheres Transportation Ambient temperature |
| title | Generation of UCiPSC-derived neurospheres for cell therapy and its application |
| title_full | Generation of UCiPSC-derived neurospheres for cell therapy and its application |
| title_fullStr | Generation of UCiPSC-derived neurospheres for cell therapy and its application |
| title_full_unstemmed | Generation of UCiPSC-derived neurospheres for cell therapy and its application |
| title_short | Generation of UCiPSC-derived neurospheres for cell therapy and its application |
| title_sort | generation of ucipsc derived neurospheres for cell therapy and its application |
| topic | Human induced pluripotent stem cells hiPSC-derived human neural stem cells Neurospheres Transportation Ambient temperature |
| url | https://doi.org/10.1186/s13287-021-02238-4 |
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