Cerebrospinal fluid used as culture medium prior to autologous olfactory ensheathing cell transplantation

作     者：Weijiang Wu Qing Lan Hua Lu Aihua Zhu Yunzhao Jiang Ge Chen Guozhen Hui 

作者机构：Department of Neurosurgery Wuxi Third People's Hospital Wuxi 214041 Jiangsu Province China Second Affiliated HospitaI Soochow University Suzhou 215004 Jiangsu Province China First Affiliated Hospital Soochow University Suzhou 215006 Jiangsu Province China 

出 版 物：《Neural Regeneration Research》 (中国神经再生研究（英文版）)

年 卷 期：2010年第5卷第1期

页      面：21-27页

核心收录：

学科分类：0710[理学-生物学] 1002[医学-临床医学] 1001[医学-基础医学(可授医学、理学学位)] 10[医学] 

基　　金：the National Trauma Program (973 Program)  No. 2005CB522600 

主　　题：cerebrospinal fluid olfactory ensheathing cells in vitro culture cell transplantation 

摘      要：BACKGROUND： Cerebrospinal fluid can be an inducer for neural stem cells in vitro, but few studies employ cerebrospinal fluid to culture olfactory ensheathing cells. OBJECTIVE： To investigate the growth of nasal mucosa olfactory ensheathing cells in normal cerebrospinal fluid, and to analyze the feasibility of cerebrospinal fluid for culturing olfactory ensheathing cells used for transplantation. DESIGN, TIME AND SETTING： A completely randomized, block design study was performed at the Cell Laboratory, Wuxi Third People＇s Hospital, and Jiangsu Institute of Parasitic Diseases, China, in August 2008. MATERIALS： Dulbecco＇s modified Eagle＇s medium/F12 （DMEM/F12） and fetal bovine serum （Gibco BRL, USA）, mouse anti-rat P75 monoclonal antibody and rabbit anti-glial fibrillary acidic protein polyclonal anti body （＇Santa Cruz Biotechnology, USA）, mouse anti-rat myelin basic protein monoclonal antibody （Cymbus, UK）, mouse anti-rat microtubule-associated protein-2 monoclonal antibody （Transduction Laboratories, USA）, FITC conjugated rabbit anti-mouse monoclonal antibody （Boster, China）, TRITC conjugated goat anti-rabbit monoclonal antibody （Sigma, USA） were used. METHODS： Nasal mucosa olfactory ensheathing cells were separately incubated in DMEM/F12, cerebrospinal fluid, and changing DMEM/F12 into cerebrospinal fluid. Adult female Sprague Dawley rat models of spinal hemisection were established. Nerve injury was repaired by transplantation of nasal mucosa olfactory ensheathing cells cultured in cerebrospinal fluid or DMEM/F12. MAIN OUTCOME MEASURES： The proliferative ability of olfactory ensheathing cells cultured in cerebrospinal fluid was determined by a 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyl tetrazolium bromide assay. The morphology and purity of olfactory ensheathing cells were detected using immunohistochemistry. Animal behavior was evaluated by the Basso, Beattie and Bresnahan locomotor rating scale. Morphological repair was assessed by a horseradish peroxidase-tetra