Int. J. Dev. Biol. In Press
doi: 10.1387/ijdb.210062zz
© UPV/EHU Press

SV40T reprograms Schwann cells into stem-like cells that can re-differentiate into terminal nerve cells

Rui-fang Li1, Guo-xin Nan2, Dan Wang1, Chang Gao1, Juan Yang1, Tong-chuan He*,3, Zhong-lin Zhang*,4

1Department of Neurology, Hubei Third People's Hospital of Jianghan University, Wuhan, China, 2Department II of Orthopaedics, Chongqing Key Laboratory of Pediatrics, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders, Children’s Hospital of Chongqing Medical University, Chongqing, China, 3Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center, Chicago, USA and 4Department of Hepatobiliary Surgery, the Research Center of Digestive Diseases, Zhongnan Hospital of Wuhan University, Wuhan, China

ABSTRACT

Background: The specific effect of SV40T on neurocytes has been rarely investigated by the researchers. We transfected Schwann cells (SCs) that did not have differentiation ability with MPH 86 plasmid containing SV40T in order to explore the effects of SV40T on Schwann cells.

Methods: SCs were transfected with MPH 86 plasmid carrying the SV40T gene and cultured in different media, as well as co-cultured with neural stem cells (NSCs). In our study, SCs overexpressing SV40T were defined as SV40T-SCs. The proliferation of these cells was detected by WST-1, and the expression of different biomarkers was analyzed by qPCR and immunohistochemistry.

Results: SV40T induced the characteristics of NSCs, such as the ability to grow in suspension, form spheroid colonies and proliferate rapidly, in the SCs, which were reversed by knocking out SV40T by the Flip-adenovirus. In addition, SV40T upregulated the expressions of neural crest-associated markers Nestin, Pax3 and Slug, and down-regulated S100b as well as the markers of mature SCs MBP, GFAP and Olig1/2. These cells also expressed NSC markers like Nestin, Sox2, CD133 and SSEA-1, as well as early development markers of embryonic stem cells (ESCs) like BMP4, c-Myc, OCT4 and Gbx2. Co-culturing with NSCs induced differentiation of the SV40T-SCs into neuronal and glial cells.

Conclusions: SV40T reprograms Schwann cells to stem-like cells at the stage of neural crest cells (NCCs) that can differentiate to neurocytes.

Keywords:

Schwann cells, SV40T, cell transformation, cell reprogramming, inverse differentiation, neural crest cells

*Corresponding author e-mail: tche@uchicago.edu ; zhonglinzhang@whu.edu.cn