目的 研究人乳牙牙髓干细胞(SHED)表达和分泌胶质细胞源性神经营养因子(GDNF)的能力及规律,为进一步探讨SHED治疗帕金森病的作用机制提供依据.方法 通过酶消化法体外分离培养人乳牙牙髓中的干细胞,当传代至第3代时,加入神经干细胞的特殊培养基Neurobasal A和细胞因子进行神经球诱导培养,通过Real-Time PCR、免疫荧光和ELISA等方法,分别从mRNA和蛋白水平检测体外培养SHED和神经球中GDNF的表达.结果 Real-Time PCR检测结果显示,在mRNA水平,SHED和神经球中都有GDNF的表达.在蛋白水平,免疫荧光结果可见SHED和神经球中都有GDNF的表达;采用ELISA法在培养第3天的SHED培养液和神经球的培养上清中均可检测到GDNF,并随着时间的延长,培养液中GDNF浓度明显升高.结论 SHED具有表达和分泌GDNF的能力.%Objective To investigate the capability of stem cells from human exfoliated deciduous teeth (SHED) to produce and secrete glial cell line-derived neurotrophic factor (GDNF). Methods SHED were isolated from the pulp of normal exfoliated human deciduous incisors by enzymatic digestion and passage in vitro. At passage 3, SHED were induced by a medium optimized for neural stem cells growth which contained Neurobasal A and supplemented with other cell growth factors for neural differentiation. The expression of GDNF was detected by real time-PCR,immunofluorescence staining and ELISA. Results The mRNA and protein of GDNF could be detected in cells and culture supernatant after 3-day culture. The expression of GDNF in culture supernatant increased gradually with time of culture. Conclusion SHED have the potential to produce and secrete GDNF.
展开▼
