Enhanced neural differentiation of neural stem cells by sustained release of Shh from TG2 gene-modified EMSC co-culture
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ORIGINAL ARTICLE
Enhanced neural differentiation of neural stem cells by sustained release of Shh from TG2 gene‑modified EMSC co‑culture in vitro Wentao Shi1 · Lu Bian1 · Demin Lv4 · Shiqi Bi4 · Yao Dai4 · Kaiyuan Yang4 · Hao Lu4 · Huangao Zhou4 · Yunduan Que3 · Dongming Wang3 · Zhijian Zhang4 · Naiyan Lu2 Received: 6 July 2020 / Accepted: 13 November 2020 © Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract As a promising cell therapy, neural crest-derived ectoderm mesenchymal stem cells (EMSCs) secrete high amounts of extracellular matrix (ECM) and neurotrophic factors, promoting neural stem cell (NSC) differentiation into neuronal lineages and aiding tissue regeneration. Additionally, the forced overexpression of secreted proteins can increase the therapeutic efficacy of the secretome. Tissue transglutaminase (TG2) is a ubiquitously expressed member of the transglutaminase family of calcium-dependent crosslinking enzymes, which can stabilize the ECM, inducing smart or living biomaterial to stimulate differentiation and enhance the neurogenesis of NSCs. In this study, we examined the neuronal differentiation of NSCs induced by TG2 gene-modified EMSCs (TG2-EMSCs) in a co-culture model directly. Two weeks after initiating differentiation, levels of the neuronal markers, tubulin beta 3 class III and growth-associated protein 43, were higher in NSCs in the TG2-EMSC co-culture group and those of the astrocytic marker glial fibrillary acidic protein were lower, compared with the control group. These results were confirmed by immunofluorescence, and laminin, fibronectin and sonic hedgehog (Shh) contributed to this effect. The results of western blot analysis and the enzyme-linked immunoassay showed that after TG2-EMSCs were co-cultured for 2 weeks, they expressed much higher levels of Shh than the control group. Moreover, the sustained release of Shh was observed in the TG2-EMSC co-culture group. Overall, our findings indicate that EMSCs can induce the differentiation of NSCs, of which TG2-EMSCs can promote the differentiation of NSCs compared with EMSCs. Keywords Tissue transglutaminase 2 · Shh · EMSC · NSC · Differentiation
Introduction Handling Editor: T. Harkany. Wentao Shi, Lu Bian and Demin Lv contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00726-020-02918-0) contains supplementary material, which is available to authorized users. * Naiyan Lu [email protected] 1
School of Medicine, Jiangnan University, Wuxi, Jiangsu 214122, People’s Republic of China
2
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, People’s Republic of China
3
Nanjing Gaochun People’s Hospital, Nanjing 211300, People’s Republic of China
4
School of Medicine, Jiangsu University, Zhenjiang, Jiangsu 212001, People’s Republic of China
Spinal cord injury (SCI) has one of the highest rates of morbidity and disability, leading to a heavy economic burden on society (Rabinstein
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