The Rotary Cell Culture System increases NTRK3 expression and promotes neuronal differentiation and migratory ability of neural stem cells cultured on collagen sponge

• Published in: Stem cell research & therapy 2021-05, Vol.12 (1), p.298-298, Article 298
• Main Authors: Cui, Yi, Yin, Yanyun, Zou, Yunlong, Zhao, Yannan, Han, Jin, Xu, Bai, Chen, Bing, Xiao, Zhifeng, Song, Hongwei, Shi, Ya, Xue, Weiwei, Ma, Xu, Dai, Jianwu
• Format: Article
• Language: English
• Subjects: Alzheimer's disease; Animals; Binding sites; Care and treatment; Cell culture; Cell Culture Techniques; Cell Differentiation; Cells, Cultured; Collagen; Collagen sponge scaffold; Environmental engineering; Enzymes; Growth factors; Health aspects; Immunofluorescence; Kinases; Laboratory animals; let-7i-5p; Microgravity; MicroRNA; miRNA; mRNA; Nervous system; Nervous system diseases; Neural Stem Cells; Neurological diseases; Neurons; Neurotrophin 3; NSCs; NTRK3; Protein-tyrosine kinase receptors; Public health; RCCS; Receptor, trkC; Spinal cord; Spinal cord injuries; Sponge culture; Stem cell transplantation; Stem cells; Tissue culture; Tissue engineering; Tissue Scaffolds; Transplantation; Tyrosine
• ISSN: 1757-6512; 1757-6512
• DOI: 10.1186/s13287-021-02381-y

Recently, neural stem cell (NSC) therapy has shown promise for the treatment of many neurological diseases. Enhancing the quality of implanted cells and improving therapeutic efficacy are currently research hotspots. It has been reported that collagen sponge material provided sufficient room for cell growth in all directions and promoted the absorption of nutrients and removal of wastes. And also, the Rotary Cell Culture System (RCCS), which mimics the microgravity environment, can be used to culture cells for tissue engineering. We performed the mRNA and miRNA sequencing to elucidate the regulatory mechanism of NSCs cultured on the collagen sponge in the RCCS system. The luciferase assay and Western blot revealed a direct regulatory role between let-7i-5p and neurotrophic receptor tyrosine kinase 3 (NTRK3; also called TrkC). And then, the neural differentiation markers Tuj1 and Map2 were detected by immunofluorescence staining. In the meantime, the migratory ability of NSCs was detected both in vitro and in spinal cord injury animals. In this study, we demonstrated that the expression of NTRK3 was elevated in NSCs cultured on collagen sponge in the RCCS system. Furthermore, increased NTRK3 expression was regulated by the downregulation of let-7i-5p. Compared to traditionally cultured NSCs, the NSCs cultured on collagen sponge in the RCCS system exhibited better neuronal differentiation and migratory ability, especially in the presence of NT-3. As the biological properties and quality of transplanted cells are critical for therapeutic success, the RCCS system combined with the collagen sponge culture system shows promise for applications in clinical practice in the future.