Neuronal differentiation of cell subsets with stem cell characteristics in adult rat meningeal tissues

doi:10.3969/j.issn.2095-4344.2015.50.024

Abstract

Abstract:

BACKGROUND: Neural stem cells have the potential to differentiate into neurons and glial cells to replace the injured brain cells, so as to achieve the purpose of repairing nerve injury.
OBJECTIVE: To observe the neuronal differentiation ability of cell subsets with stem cell characteristics in the adult rat meningeal tissues.
METHODS: Under anesthesia, the meningeal tissues were obtained from adult Sprague-Dawley rats to make cell suspension followed by inoculation and subculture. Then, the Nestin immunofluorescence staining was performed. The third generation cells were cultured in vitro with complete culture medium containing trichostatin A. After 7 days of induction, western blot assay was used to detect the expression of NF-200 and BM88 proteins in neural cells.
RESULTS AND CONCLUSION: At 24 hours of culture, some spherical cells were suspended and some cells adherent. In addition, some spherical cells scattered gradually formed the clone spheres, and the growth rate decreased with the increasing volume. The positive expression of Nestin was detected by immunocytochemistry staining, and the cell nucleus was stained blue by Hoechst staining. BM88 and NF-200 proteins were all expressed at 7 days of neural induction. These findings indicate that the cell subsets with stem cell characteristics in the adult rat meningeal tissues can differentiate into neurons after in vitro induction.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

CLC Number:

R394.2

Liu De, Li Xiang-ming, Gu Xi-juan. Neuronal differentiation of cell subsets with stem cell characteristics in adult rat meningeal tissues[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(50): 8172-8176.

Figures/Tables 4

References

[1] 孙晓飞,金光辉,谢杨,等.神经干细胞移植治疗脊髓损伤的研究进展[J].中国矫形外科杂志,2015,23(18):1680-1682.

[2] 杨万章,吴芳,张敏,等.脐血源神经干细胞移植治疗神经系统疾病临床总结和分析[J].中西医结合心脑血管病杂志,2009,7(3): 287-290.

[3] 陈东,孟晓婷,刘佳梅,等.鹿茸多肽对胎大鼠脑神经干细胞体外诱导分化的实验研究[J].解剖学报,2004,35(3):240-243.

[4] Menendez L, Yatskievych TA, Antin PB, et al. Wnt signaling and a Smad pathway blockade direct the differentiation of human pluripotent stem cells to multipotent neural crest cells. Proc Natl Acad Sci U S A. 2011;108(48):19240-19245.

[5] Li YC, Liao YT, Chang HH, et al. Covalent bonding of GYIGSR to EVAL membrane surface to improve migration and adhesion of cultured neural stem/precursor cells. Colloids Surf B Biointerfaces. 2013;102:53-62.

[6] 张宝华,仇福成,董慈,等.脑脊液途径移植神经干细胞治疗中枢神经系统疾病的研究现状[J].中国组织工程研究,2014,18(6): 974-978.

[7] 吴家华,罗焕敏.神经干细胞移植治疗中枢神经系统疾病可行性研究[J].中国老年学杂志,2005,25(1):110-112.

[8] 温秀杰,刘鲁川,王尧,等.碱性成纤维生长因子和表皮生长因子对脊髓神经干细胞增殖与分化的影响[J].中国临床神经科学,2014, 22(2):121-125.

[9] 闫峰,张越林,刘宁,等.全骨髓贴壁法分离骨髓间充质干细胞并诱导其定向神经元样细胞的分化[J].细胞与分子免疫学杂志,2014, 30(11):1162-1165.

[10] 牛平,王耀山,吕永利,等.神经营养因子对体外培养多巴胺能神经元营养作用比较研究[J].中国医科大学学报,2001,30(6):408-410.

[11] 郭洪波,邹飞.大鼠海马神经干细胞的分离、培养及鉴定[J].第一军医大学学报,2004,24(10):1143-1146.

[12] 谭新杰,胡长林,蔡文琴,等.新生大鼠海马神经干细胞的分离、培养、分化和鉴定[J].国际脑血管病杂志,2006,14(2):107-109.

[13] 丁道芳,邢三丽,周鸣鸣,等.大鼠胚胎神经干细胞单克隆化及单层化培养和鉴定[J].生物化学与生物物理进展,2009,36(1):72-76.

[14] 史冬梅,周畅,谢佐平,等.神经干细胞有丝分裂过程中nestin表达变化[J].神经解剖学杂志,2003,19(2):119-123.

[15] 林旭妍,林绿标,许益民,等.神经膜细胞促进神经干细胞存活及分化的实验研究[J].中华脑科疾病与康复杂志:电子版,2012,2(5): 24-27.

[16] 杨忠旭,栾国明.神经干细胞的定向分化调控及移植治疗颞叶癫痫[J].立体定向和功能性神经外科杂志,2003,16(2):105-107.

[17] Heng BC, Saxena P, Fussenegger M. Heterogeneity of baseline neural marker expression by undifferentiated mesenchymal stem cells may be correlated to donor age. J Biotechnol. 2014;174:29-33.

[18] Caprini A, Silva D, Zanoni I, et al. A novel bioactive peptide: assessing its activity over murine neural stem cells and its potential for neural tissue engineering. N Biotechnol. 2013; 30(5):552-562.

[19] 陈娟,李娟,谭军,等.腺苷预处理对局灶性脑缺血再灌注损伤大鼠脑组织中内源性神经干细胞增殖的影响[J].郑州大学学报:医学版,2014,49(5):645-647,648.

[20] 许锋,姜红,周春清,等.促红细胞生成素对新生鼠缺氧缺血性脑损伤后神经干细胞的影响[J].中国新生儿科杂志, 2010, 25(2): 94-97.

[21] 唐蔚东,叶建亚,李彩格,等.巢蛋白和性别决定基因高迁移率组蛋白阳性细胞在成年大鼠穹隆下器的表达[J].解剖学杂志,2014, 37(2):202-204,220.

[22] 王婷.柔脑膜细胞具有神经干细胞特性的实验研究[D].西安:解放军第四军医大学,2008.

[23] Zou Z, Liu T, Li J, et al. Biocompatibility of functionalized designer self-assembling nanofiber scaffolds containing FRM motif for neural stem cells. J Biomed Mater Res A. 2014; 102(5): 1286-1293.

[24] 顾繁,高建华,鲁峰.hASCs的单克隆培养及干细胞相关标志物表达的实验研究[J].南方医科大学学报,2008,28(6):1067-1069,1075.

[25] 丛琳,朱晓峰.FGF-2,NT-3对海马表皮生长因子反应性神经干细胞分化为神经元的诱导作用[J].神经损伤与功能重建,2006, 1(1): 25-27,37.

[26] 李晓波,丁新生,潘凤华,等.碱性成纤维细胞生长因子、表皮生长因子对脑缺血大鼠内源性神经干细胞增殖的影响[J].临床神经病学杂志,2006,19(3):188-190.

[27] 段淑荣,王慧慧,戚基萍,等.脑缺血后碱性成纤维细胞生长因子、表皮生长因子对内源性神经干细胞增殖的影响[J].中华医学杂志,2008,88(47):3337-3341.

[28] 向云,燕铁斌,金冬梅,等.低频电刺激对急性脑梗死大鼠脑bFGF、EGF表达和内源性神经干细胞增殖的影响[J].中华物理医学与康复杂志,2010,32(12):881-886.

[29] Khosravizadeh Z, Razavi S, Bahramian H, et al. The beneficial effect of encapsulated human adipose-derived stem cells in alginate hydrogel on neural differentiation. J Biomed Mater Res B Appl Biomater. 2014;102(4):749-755.

[30] Lv Y, Zhao P, Chen G, et al. Effects of low-intensity pulsed ultrasound on cell viability, proliferation and neural differentiation of induced pluripotent stem cells-derived neural crest stem cells. Biotechnol Lett. 2013;35(12):2201-2212.

[31] Zhang Y, Guo Y, Wu B, et al. Ligand-mediated endocytosis of nanoparticles in neural stem cells: implications for cellular magnetic resonance imaging. Biotechnol Lett. 2013;35(12): 1997-2004.

[32] 陈东风,杜少辉,李伊为,等.龟板对局灶性脑缺血再灌注后Nestin表达的影响[J].解剖学杂志,2002,25(4):315-319.

[33] 程龙,朱培纯,司银楚,等.三七总皂甙对去大脑皮层血管后成年大鼠前脑侧脑室室管膜下层Nestin和bFGF表达的作用[J].北京中医药大学学报,2003,26(3):18-20.

[34] 王军,范亚珍,陈海,等.脐血单个核细胞诱导的神经干细胞中Foxg1和Nestin基因的表达及其相互关系[J].实用儿科临床杂志,2010,25(11):848-850,868.

[35] Ravindran G, Devaraj H.Prognostic significance of neural stem cell markers, Nestin and Musashi-1, in oral squamous cell carcinoma: expression pattern of Nestin in the precancerous stages of oral squamous epithelium.Clin Oral Investig. 2015;19(6):1251-1260.

[36] Di CG, Xiang AP, Jia L,et al. Involvement of extracellular factors in maintaining self-renewal of neural stem cell by nestin. Neuroreport. 2014;25(10):782-787.

[37] Miao HC, Wu F, Ding J, et al. Effects of electroacupuncture intervention combined with polysaccharide of Gastrodia elata Blume on expression of nestin and cytokines of neural stem cells in the dentate gyrus of cerebral ischemia rats. Zhen Ci Yan Jiu. 2014;39(1):40-45.

[38] Gao WL, Tian F, Zhang SQ, et al.Epidermal growth factor increases the expression of Nestin in rat reactive astrocytes through the Ras-Raf-ERK pathway. Neurosci Lett. 2014; 562:54-59.

[39] Tomita T, Akimoto J, Haraoka J, et al. Clinicopathological significance of expression of nestin, a neural stem/progenitor cell marker, in human glioma tissue. Brain Tumor Pathol. 2014;31(3):162-171.

[40] Yang P, Jin H, Xiao X,et al.Ventricular and subventricular zones under the frontal cortex of human fetus: development and distribution of nestin-positive cells.Nan Fang Yi Ke Da Xue Xue Bao. 2013;33(5):708-714.

[41] Wu MD, Montgomery SL, Rivera-Escalera F, et al.Sustained IL-1β expression impairs adult hippocampal neurogenesis independent of IL-1 signaling in nestin+ neural precursor cells. Brain Behav Immun. 2013;32:9-18.

[42] Shin YJ, Kim HL, Park JM, et al. Characterization of nestin expression and vessel association in the ischemic core following focal cerebral ischemia in rats. Cell Tissue Res. 2013;351(3):383-395.