CAJ | 학술논문

目的研究成人脂肪基质细胞(ADSC)诱导分化为神经前体细胞数量达峰时间和诱导分化后神经前体细胞的超微结构特征.方法应用β-巯基乙醇诱导成人ADSC向神经前体细胞和神经元样细胞分化,倒置相差显微镜观察诱导前后细胞形态;免疫荧光细胞化学染色法检测诱导分化后细胞神经巢蛋白(nestin)的表达情况;透射电镜观察诱导分化3h时的神经前体细胞超微结构.结果体外培养的成人ADSC诱导分化3h时nestin的表达到高峰,为(86.25±4.82)%,透射电镜观察神经前体细胞的超微结构可见细胞表面突起较多,胞质中细胞器丰富,细胞核大,核/浆比例大,双层核膜且有较多核孔,常染色质多,异染色质少.结论 β-巯基乙醇诱导成人ADSC分化3h时神经前体细胞数量达到高峰,且超微结构研究证实此时细胞处于分裂增殖旺盛期.
목적 연구성인지방기질세포(ADSC)유도분화위신경전체세포수량체봉시간화유도분화후신경전체세포적초미결구특정.방법 응용β-구기을순유도성인ADSC향신경전체세포화신경원양세포분화,도치상차현미경관찰유도전후세포형태;면역형광세포화학염색법검측유도분화후세포신경소단백(nestin)적표체정황;투사전경관찰유도분화3h시적신경전체세포초미결구.결과 체외배양적성인ADSC유도분화3h시nestin적표체도고봉,위(86.25±4.82)%,투사전경관찰신경전체세포적초미결구가견세포표면돌기교다,포질중세포기봉부,세포핵대,핵/장비례대,쌍층핵막차유교다핵공,상염색질다,이염색질소.결론 β-구기을순유도성인ADSC분화3h시신경전체세포수량체도고봉,차초미결구연구증실차시세포처우분렬증식왕성기.
Objective To reseach the time point of the highest percentage of neural precursor cells derived from adipose stromal cells (ADSCs) in vitro, and to observe the ultrastructure features of neural precursor cells. Methods Used the β-mercaptoethanol to induce ADSCs to differentiate into neural precursor cells and neuron-like cells. The morphology of the uninductedcells and inducted cells were observed with inverted phase contrast microscope. The expression of nestin which was the marker of neural precursor cell in each group was detected using immunofluorescence staining method. The ultrastructural feature of cells which was induced for 3 hours were observed. Results The highest ratio of positive expression of nestin was 3 hours following induction,with the ratio ( 86.25 ± 4.82) %. There were many protuberance on the cell membrane under transmission electron microscopy.There were plenty of organelles in the neural precursor cells. The neural precursor cells had a large size nucleus,large nucleoplasmic index, much extended chromatin,and less condensed chromatin. The nucleus had double-layer nuclear envelope, more nuclear pore on the nuclear envelope. Conclusion The time point of the highest percentage of neural precursor cells derived from ADSCs is 3 hours,and the ultrastructral feature of induced neural precursor cells confirm that cells at this time point are in a state of split active period.