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目的 探讨转录因子Brn 3a诱导胚胎干细胞 (ES细胞 )向神经细胞定向分化的可能性。 方法 将带有目的基因 (Brn 3a转录因子 )的重组表达载体pJ5Brn 3a ,通过脂质体转染到小鼠ES细胞株中进行表达 ,促使ES细胞向神经细胞分化 ,并采用免疫组织化学技术检测转染前后转录因子Brn 3a蛋白及分化后具有神经元表型特征的神经样细胞特异抗原的表达。 结果 1 转染前ES细胞Brn 3a免疫反应阴性 ,转染后 2 4h检测Brn 3a免疫反应呈阳性 ;2 转染细胞经 1周培养 ,70 %以上的细胞具有明显的神经细胞样突起 ,神经细胞特有标记抗原NFH +L、NSE及SY呈免疫反应阳性 ;神经胶质细胞特有标记抗原GFAP为阴性。 结论 转录因子Brn 3a能成功诱导ES细胞向神经细胞定向分化
Objective To investigate the possibility of directional differentiation of embryonic stem cells (ES cells) into neurons by transcription factor Brn 3a. Methods The recombinant plasmid pJ5Brn 3a carrying the target gene (Brn 3a transcription factor) was transfected into mouse ES cell line by lipofectamine to express ES cells and induce the differentiation of ES cells into neurons. The expression of pJ5Brn 3a was detected by immunohistochemistry Expression of Brn 3a transcription factor before and after transfection and expression of neuron - like cell specific antigen with neuronal phenotype after differentiation. Results 1 The Brn 3a immunoreactivity of ES cells before transfection was negative, and the Brn 3a immunoreactivity was detected at 24 hrs after transfection. 2 After 1 week of transfection, more than 70% of the cells had obvious neuron-like processes, neurons The specific marker antigen NFH + L, NSE and SY were immunoreactive; GFAP marker-specific glial cells was negative. Conclusion Transcription factor Brn 3a can successfully induce ES cells to differentiate into neurons