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

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

doi:10.3892/etm.2020.9285

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

Exp Ther Med. 2020 Dec;20(6):156. doi: 10.3892/etm.2020.9285. Epub 2020 Oct 6.

Authors

Hengxing Zhou^{1

2}, Huan Yang³, Lu Lu^{1

2}, Xueying Li⁴, Bin Pan^{1

2}, Zheng Fu⁴, Tianci Chu⁵, Jun Liu^{1

2}, Yi Kang^{1

2}, Lu Liu^{1

2}, Guangzhi Ning^{1

2}, Wenyuan Ding⁶, Ping Wu⁷, Xiaohong Kong⁸, Shiqing Feng^{1

2}

Affiliations

¹ Department of Orthopedics, Tianjin Medical University General Hospital, Tianjin 300052, P.R. China.
² Key Laboratory of Post-Neuroinjury Neuro-Repair and Regeneration in Central Nervous System, Tianjin Neurological Institute, Ministry of Education and Tianjin City, Tianjin 300052, P.R. China.
³ Department of Orthopedics, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu 221006, P.R. China.
⁴ Key Laboratory of Immuno Microenvironment and Disease of the Educational Ministry of China, Department of Immunology, Tianjin Medical University, Tianjin 300070, P.R. China.
⁵ Department of Pediatrics, Kosair Children's Hospital Research Institute, University of Louisville School of Medicine, Louisville, KY 40202, USA.
⁶ Department of Orthopedics, The Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China.
⁷ Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, TX 77555-1043, USA.
⁸ School of Medicine, Nankai University, Tianjin 300071, P.R. China.

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).

Keywords: cryopreservation; multipotency; neural stem cells; primary culture; self-renewal.