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| 1 | In vitro culture and differentiation of rat embryonic midbrain-derived neural stem cells显示文摘BACKGROUND:Midbrain-derived neural stem cells (mNSCs) can differentiate into functional mature dopaminergic neurons. The mNSCs are considered the ideal choice for cell therapy of Parkinson’s disease. OBJECTIVE: To isolate rat embryonic mNSCs and to observe the differentiation characteristics of mNSCs induced by cell growth-promoting factors. DESIGN,TIME AND SETTING: An in vitro cell culture study based on the molecular biology of nerve cells was carried out at the Institute of Clinical Medicine,China-Japan Friendship Hospital (China) from March to November 2007. MATERIALS: Sprague Dawley rats at embryonic day 14 were used in this study. Nestin antibody,β-Ⅲ tubulin antibody,glial fibrillary acidic protein (GFAP) antibody and cyclic nucleotide 3’-phosphohydrolase (CNPase) antibody were provided by Abcam; DMEM/F12 medium and N2 supplement were provided by In-vitrogen; epidermal growth factor (EGF) and fibroblast growth factor-2 (FGF2) were provided by R&D Sys-tems. METHODS: The ventral mesencephalon was dissected from embryonic day 14 rat embryos. By trypsin di-gestion and mechanical separation,the brain tissue was triturated into a fine single-cell suspension. The cells were cultured in 5 mL serum-free medium containing DMEM/F12,1% N2 supplement,20 ng/mL EGF and FGF2. The mNSCs at the third generation were coated with 10 μg/mL polylysine and induced to differenti-ate in the DMEM/F12 supplemented with 1% fetal bovine serum and 1% N2. MAIN OUTCOME MEASURES: The neural spheres of the third passage were identified by nestin im-munofluorescence; at the same time,the cells were induced to differentiate,and the types of differentiated cell were identified by immunofluorescence for βⅢ tubulin,GFAP and CNPase. RESULTS: Seven days after primary culture,a great many neurospheres could be obtained by successive pasage. Immunofluorescence assays showed that the neurospheres were nestin positive,and after differentia-tion,the cells expressed GFAP,CNPase and β-Ⅲ-tubulin. CONCLUSION: Embryonic day 14 rat mNSCs can differentiate into neuron-like cells and glial cells fol-lowing induction by EGF,FGF2 and N2 additive. | Xingli Deng Ruen Liu Zhongtang Feng Jing Guo Wu Wang Deqiang Lei Hongyan Li Zhihua Chen | 2008 | Neural Regeneration Research2008，3，11： | 19 |
| 2 | Long-term follow-up of cancer patients receiving radiotherapy for bone metastases:results from a randomised multicentre trial显示文摘 | Sande TA Ruenes R Lund JA | | 0，，02： | 1 |
| 3 | Neuronal damage following transpinal injection of a nitric oxide desynthase inhibitor in the rat显示文摘 | YEZIERSKI RP LIU S RUENES G L | 1996 | J Cereb Blood Flow Metab1996，16，5： | 1 |
| 4 | A distinct dominant form of mierotia and conductive hearing loss显示文摘 | Sanchez -Corona J Garcia -Cruz D Ruenes R | 1982 | Birth Defects Orig Artic Ser1982，18，3： | 1 |
| 5 | Plasmacytoma of the pet-rous temporal bone ear显示文摘 | RUENES R PALACIOS E | 2003 | Nose Throat J2003，82，： | 1 |
| 6 | A distinct dominant form of mierotia and conductive hearing loss显示文摘 | Sanches-Corona J Garcia-Cruz D Ruenes R | 1982 | Birth Defects Orig Attic Ser1982，18，3： | 1 |
| 7 | Excitotoxic spinal cord injury:behavioral and morphological characteristics of a central pain model显示文摘 | Yezierski RP Liu S Ruenes GL | 1998 | Pain1998，75，1： | 1 |
| 8 | Application of KBE technology in Die & Mold Design显示文摘 | Peng Ying-hong Zhao Zhen Ruen Xue-yu | 2000 | International Conference on Die end Mold Technology Beijing2000，9086，： | 1 |
| 9 | Malignant fibrous histiocytoma of the small bowel with colon metastasis显示文摘 | Villa Jimenez OM Herngndez Garces HR Ruenes Domech C | 2011 | Endoscopy2011，43，2： | 1 |
| 10 | Neuronal damage following intraspinal injection of a nitric oxide synthase in hibitor inthe rat 显示文摘 | Yezierski RP Liu S Ruenes GL | 1996 | J Cereb Blood Flow Metab1996，16，5： | 1 |
| 11 | Utilization of bedrock water by Brosimum alicastrum trees growing on shallow soil atop limestone in a dry tropical climate显示文摘 | José Ignacio Querejeta Héctor Estrada-Medina Michael F. Allen Juan J. Jiménez-Osornio Rocío Ruenes | 2006 | Plant and Soil (-)2006，，1： | 1 |
| 12 | Dptimal local weighted averaging methods in contour smoothing显示文摘 | Legauit R Ruen C Y | 1997 | IEEE Trans PAMI1997，19，9： | 1 |
| 13 | Excitotoxic spinal cord injury:behavioral and morphological characteristics of a central pain model显示文摘 | Yezierski RP Liu S Ruenes GL | 1998 | Pain1998，75，1： | 1 |
| 14 | Neuronal damage following intraspinal injection of a nitric oxide synthase inhibitor in the rat 显示文摘 | Yezierski RP Liu S Ruenes GL | 1996 | Cereb Blood Flow Metab1996，16，5： | 1 |
| 15 | Long-term follow-up of cancer patients receiving radiotherapy for bone metastases: Results from a randomised multicentre trial显示文摘 | Tonje Anette Sande Randi Ruenes Jo Asmund Lund Oyvind S. Bruland Kjersti Hornslien Roy Bremnes Stein Kaasa | 2009 | Radiotherapy and Oncology2009，，2： | 1 |
| 16 | Conditions affecting the onset, severity, and progression of a spontaneous pain-like behavior after excitotoxic spinal cord injury显示文摘 | Gorman AL Yu CG Ruenes GR | 2001 | J Pain2001，2，4： | 1 |
| 17 | Modelling stream - aquifer seepage in an alluvial aquifer:an improved loosing- stream pack- age for MODFLOW显示文摘 | Yassin Z Osman Michael P ruen | 2002 | Journal of Hydrology2002，264，： | 1 |
| 18 | Plasmacytoma of the petrous temporal bone Ear显示文摘 | Ramon Ruenes Enrique Palacios | 2003 | Nose & Throat Journal2003，82，： | 1 |
| 19 | Cyanamide activation of peroxide bleaching:a lignin model compound study显示文摘 | Ruen Smit Ian D Suckling Richard M EDE | 2000 | Tappi2000，，6： | 1 |
| 20 | Increase in a Dutch hospital of methicillin-resistantStaphylococcus aureus related to animal farming 显示文摘 | VAN RUEN M M L VAN KEULEN P H KLUYTMANSJ A | 2008 | Clin-Infect Dis2008，46，2： | 1 |
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