Effect of serum on the differentiation of neural stem cells_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

LIU Qingxi ^1,2 , LÜ Lihui ¹ , SUN He ² , ZHANG Jinhua ² ,  MA Wenjian ^1,3 ,  ZHANG Tongcun ¹

1. College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, P.R.China;
2. Tianjin Weikai Bioeng Ltd., Tianjin, 300457, P.R.China;
3. College of Biotechnology, Qilu Institute of Technology, Jinan Shandong, 250200, P.R.China;

Corresponding author：

MA Wenjian, Email: ma_wj@tust.edu.cn; ZHANG Tongcun, Email: tony@tust.edu.cn

Keywords：

Neural stem cell; serum; cell differentiation; glial cells; neurons; rat

DOI：

10.7507/1002-1892.201710113

Video：

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ObjectiveTo investigate the effect of serum on the differentiation of neural stem cells.MethodsThe neural stem cells were isolated from the embryonic hippocampus tissues of Sprague Dawley rats at 14 day of pregnancy. After culturing and passaging, the 3rd generation cells were identified by immunocytochemical staining. Then, the cells were divided into 3 groups according to the concentrations of fetal bovine serum (FBS) used in the differentiation cell culture medium: 5% (group A), 1% (group B), 0 (group C), respectively. The other components of the culture media in 3 groups were the same. Cell viability was determined by using the Live/Dead cell staining at 8 days; the expressions of glial cell marker [glial fibrillary acidic protein (GFAP)] and neuronal marker (β-Ⅲ Tubulin) were determined and analyzed by immunocytochemical staining and real-time fluorescent PCR at 4 and 8 days of culture.ResultsBased on cell morphology and immunocytochemical staining, neural stem cells were identified. Cells were growing well with no death in all groups. With decreasing FBS concentration, the expression of GFAP was significantly decreased on both protein and mRNA level, whereas the expression of β-Ⅲ Tubulin was evidently increased. The staining of each group at 8 days was more obvious than that at 4 days. There were significant differences in mRNA expressions of GFAP and β-Ⅲ Tubulin at 4 and 8 days between groups (P<0.05).ConclusionSerum can promote the differentiation of neural stem cells into glial cells. At the same time, it inhibits the differentiation of neural stem cells into neurons, the lower the serum concentration, the smaller the effect.

Citation： LIU Qingxi, LÜ Lihui, SUN He, ZHANG Jinhua, MA Wenjian, ZHANG Tongcun. Effect of serum on the differentiation of neural stem cells. Chinese Journal of Reparative and Reconstructive Surgery, 2018, 32(2): 223-227. doi: 10.7507/1002-1892.201710113 Copy

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2.	Reynolds BA, Weiss S. Clonal and population analyses demonstrate that an EGF-responsive mammalian embryonic CNS precursor is a stem cell. Biol, 1996, 175(1): 1-13.
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14.	Bansod S, Kageyama R, Ohtsuka T. Hes5 regulates the transition timing of neurogenesis and gliogenesis in mammalian neocortical development. Development, 2017, 144(17): 3156-3167.
15.	Zhang W, Gu GJ, Zhang Q, et al. NSCs promote hippocampal neurogenesis, metabolic changes and synaptogenesis in APP/PS1 transgenic mice. Hippocampus, 2017, 27(12): 1250-1263.
16.	Wang Y, Ma J, Li N, et al. Microfluidic engineering of neural stem cell niches for fate determination. Biomicrofluidics, 2017, 11(1): 014106.
17.	陈珊珊, 彭敏, 周国忠, 等. 胎牛血清对大鼠胚胎海马神经干细胞向 GFAP 阳性细胞分化的影响. 中风与神经疾病杂志, 2014, 31(12): 1060-1063.
18.	范广明, 张文彬, 张赛, 等. 神经生长因子联合神经干细胞移植治疗大鼠脊髓损伤. 中国组织工程研究与临床康复, 2010, 14(14): 2572-2578.
19.	Zhu Z, Gorman MJ, McKenzie LD, et al. Zika virus has oncolytic activity against glioblastoma stem cells. J Exp Med, 2017, 214(10): 2843-2857.
20.	Choi KA, Choi Y, Hong S. Stem cell transplantation for Huntington’s diseases. Methods, 2017. DOI: 10.1016/j.ymeth.2017.08.017.
21.	Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assay. J Immunol Methods, 1983, 65(1-2): 55-63.
22.	张宝红, 张为宇, 韩忠燕, 等. 牛血清中蛋白组分初步分析及其对细胞培养的影响. 动物医学进展, 2009, 30(1): 33-37.

1. Moore L, Skop NB, Rothbard DE, et al. Tethered growth factors on biocompatible scaffolds improve stemness of cultured rat and human neural stem cells and growth of oligodendrocyte progenitors. Methods, 2017. [Epub ahead of print].
2. Reynolds BA, Weiss S. Clonal and population analyses demonstrate that an EGF-responsive mammalian embryonic CNS precursor is a stem cell. Biol, 1996, 175(1): 1-13.
3. Annese T, Corsi P, Ruggieri S, et al. Isolation and characterization of neural stem cells from dystrophic mdx mouse. Exp Cell Res, 2016, 343(2): 190-207.
4. 王立君, 姚晓辉. 神经递质与神经干细胞分化. 中国组织工程研究与临床康复, 2008, 12(25): 4917-4920.
5. 黄强. 胶质瘤干细胞与神经干细胞研究进展. 实用肿瘤杂志, 2004, 19(4): 267-269.
6. 王晓静, 张更林, 张衡. 神经干细胞的研究进展. 神经解剖学杂志, 2005, 21(3): 323-328.
7. Maric D, Maric I, Chang YH, et al. Prospective cell sorting of embryonic rat neural stem cells and neuronal and glial progenitors reveals selective effects of basic fibroblast growth factor and epidermal growth factor on self-renewal and differentiation. J Neurosci, 2003, 23(1): 240-245.
8. Tchieu J, Zimmer B, Fattahi F, et al. A modular platform for differentiation of human PSCs into all major ectodermal lineages. Cell Stem Cell, 2017, 21(3): 399-410.e7.
9. Pla P, Moore R, Morali OG, et al. Cadherins in neural crest cell development and transformation. Cell Physiol, 2001, 189(2): 121-132.
10. 刘靖, 曲静, 徐晓静, 等. 神经干细胞迁移的研究进展. 中国细胞生物学学报, 2012, 34(3): 201-211.
11. Yu Z, Men Y, Dong P. Schwann cells promote the capability of neural stem cells to differentiate into neurons and secret neurotrophic factors. Exp Ther Med, 2017, 13(5): 2029-2035.
12. Jiao S, Liu Y, Yao Y, et al. miR-124 promotes proliferation and differentiation of neuronal stem cells through inactivating Notch pathway. Cell Biosci, 2017, 7: 68.
13. 郑衍芳, 李艳君, 钟震亚, 等. 冬虫夏草含药血清诱导神经干细胞定向分化的影响. 时珍国医国药, 2014, 25(3): 576-578.
14. Bansod S, Kageyama R, Ohtsuka T. Hes5 regulates the transition timing of neurogenesis and gliogenesis in mammalian neocortical development. Development, 2017, 144(17): 3156-3167.
15. Zhang W, Gu GJ, Zhang Q, et al. NSCs promote hippocampal neurogenesis, metabolic changes and synaptogenesis in APP/PS1 transgenic mice. Hippocampus, 2017, 27(12): 1250-1263.
16. Wang Y, Ma J, Li N, et al. Microfluidic engineering of neural stem cell niches for fate determination. Biomicrofluidics, 2017, 11(1): 014106.
17. 陈珊珊, 彭敏, 周国忠, 等. 胎牛血清对大鼠胚胎海马神经干细胞向 GFAP 阳性细胞分化的影响. 中风与神经疾病杂志, 2014, 31(12): 1060-1063.
18. 范广明, 张文彬, 张赛, 等. 神经生长因子联合神经干细胞移植治疗大鼠脊髓损伤. 中国组织工程研究与临床康复, 2010, 14(14): 2572-2578.
19. Zhu Z, Gorman MJ, McKenzie LD, et al. Zika virus has oncolytic activity against glioblastoma stem cells. J Exp Med, 2017, 214(10): 2843-2857.
20. Choi KA, Choi Y, Hong S. Stem cell transplantation for Huntington’s diseases. Methods, 2017. DOI: 10.1016/j.ymeth.2017.08.017.
21. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assay. J Immunol Methods, 1983, 65(1-2): 55-63.
22. 张宝红, 张为宇, 韩忠燕, 等. 牛血清中蛋白组分初步分析及其对细胞培养的影响. 动物医学进展, 2009, 30(1): 33-37.

Chinese Journal of Reparative and Reconstructive Surgery

Effect of serum on the differentiation of neural stem cells

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