A modiﬁed protocol for the isolation, culture, and cryopreservation of rat embryonic neural stem cells

Zhou,Hengxing; Yang,Huan; Lu,Lu; Li,Xueying; Pan,Bin; Fu,Zheng; Chu,Tianci; Liu,Jun; Kang,Yi; Liu,Lu; Ning,Guangzhi; Ding,Wenyuan; Wu,Ping; Kong,Xiaohong; Feng,Shiqing

doi:10.3892/etm.2020.9285

A modiﬁed protocol for the isolation, culture, and cryopreservation of rat embryonic neural stem cells

Authors:
- Hengxing Zhou
- Huan Yang
- Lu Lu
- Xueying Li
- Bin Pan
- Zheng Fu
- Tianci Chu
- Jun Liu
- Yi Kang
- Lu Liu
- Guangzhi Ning
- Wenyuan Ding
- Ping Wu
- Xiaohong Kong
- Shiqing Feng
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Affiliations: Department of Orthopedics, Tianjin Medical University General Hospital, Tianjin 300052, P.R. China, Department of Orthopedics, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu 221006, P.R. China, Key Laboratory of Immuno Microenvironment and Disease of the Educational Ministry of China, Department of Immunology, Tianjin Medical University, Tianjin 300070, P.R. China, Department of Pediatrics, Kosair Children's Hospital Research Institute, University of Louisville School of Medicine, Louisville, KY 40202, USA, Department of Orthopedics, The Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China, Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, TX 77555‑1043, USA, School of Medicine, Nankai University, Tianjin 300071, P.R. China
Published online on: October 6, 2020 https://doi.org/10.3892/etm.2020.9285
Article Number: 156

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Abstract

Neural stem cells (NSCs) are characterized by their potential for self‑renewal and ability to differentiate into neurons, astrocytes, and oligodendrocytes. They are of great value to scientific studies and clinical applications. Culturing NSCs in vitro is important for characterizing their properties under controlled environmental conditions that may be modified and monitored accurately. The present study explored a modified, detailed and efficient protocol for the isolation, culture and cryopreservation of rat embryonic NSCs. In particular, the viability, nestin expression, and self‑renewal and multi‑differentiation capabilities of NSCs cryopreserved for various periods of time (7 days, or 1, 6 or 12 months) were characterized and compared. Rat embryonic NSCs were successfully obtained and maintained their self‑renewal and multipotent differentiation capabilities even following long‑term cryopreservation (for up to 12 months).

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