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目的 :通过胚胎MPC体外培养获取PD患者CRT治疗所需的DA能神经元。方法 :取自E12鼠胚中脑腹侧的MPC悬液在添加bFGF的DMEM/F12 /N2培养液中原代培养 ,鉴定培养细胞的增殖和分化能力。结果 :在添加了bFGF 10ng·mol-1的DMEM/F12 /N2培养液中MPC增殖良好 ,体外培养 5～ 7d后细胞数量扩增到培养前的 9 782± 0 0 47倍 ;培养液中撤去bFGF后细胞可分化成星形胶质细胞和神经元 ,其中多数Tuj1抗原标记阳性的神经元同时呈TH抗原标记阳性 ,培养细胞分化为DA能神经元的比例约为2 4 3 4% (10 2 / 419)。结论 :E12鼠胚MPC原代培养是获取DA能神经元的可靠途径 ,类似的技术应用于临床可能缓解PD患者CRT治疗供体不足的矛盾。
OBJECTIVE: To obtain the DA neurons required for CRT therapy in PD patients by in vitro culture of embryonic MPC. Methods: The MPC suspension from the ventral midbrain of E12 mouse embryos was primary cultured in DMEM / F12 / N2 medium supplemented with bFGF to identify the proliferation and differentiation ability of cultured cells. Results: The proliferation of MPC in DMEM / F12 / N2 medium supplemented with 10 ng · mol-1 of bFGF was good, and the number of cells proliferated from 9 782 ± 0 0 47 times before culturing in 5-7 days. The culture medium was removed After bFGF, the cells differentiated into astrocytes and neurons. Most of the neurons labeled with Tuj1 antigen were positive for TH antigen. The percentage of cultured cells differentiated into DA neurons was about 224% (10 2/419). CONCLUSION: Primary culture of E12 murine embryos with MPC is a reliable way to obtain DA neurons. A similar technique may be used clinically to alleviate the contradictions of donors in PD patients undergoing CRT therapy.