CAJ | 학술논문

背景:骨髓基质细胞最终成为神经元需要经历定向及分化两个过程,定向及分化是含有相同基因库的细胞不同基因表达的结果,基因表达需要一定的条件,胞外基质的变化可引起细胞形态学及基因表达方式的改变.目的:观察骨髓基质细胞在损伤大鼠脑组织提取液诱导下,向神经元样细胞分化的可能性.方法:取第5代转染绿色荧光蛋白的骨髓基质细胞,分别用缺血再灌注/正常大鼠脑组织提取液进行诱导分化培养,并设立空白对照.相差显微镜下观察细胞形态变化,并行免疫组织化学染色鉴定.结果与结论:原代培养的骨髓基质细胞纯化、扩增后呈均匀一致的长梭形,第3代细胞均一表达CD44,CD106,不表达CD34.荧光显微镜下,绿色荧光蛋白转染后24 h可观察到骨髓基质细胞有荧光表达,但强度稍弱;48 h后大多数细胞发出明显绿色荧光.加入缺血再灌注大鼠脑组织提取液后,诱导细胞不仅在形态上表现为神经元样特征,而且神经元特异性烯醇化酶特异性抗体呈阳性表达.与空白对照组比较,缺血再灌注大鼠脑组织提取液组和正常大鼠脑组织提取液组骨髓基质细胞分化率均明显升高(P < 0.05),且前组升高幅度明显大于后组(P < 0.05).提示缺血再灌注大鼠脑组织提取液能将骨髓基质细胞成功诱导为神经元样细胞.
배경:골수기질세포최종성위신경원수요경력정향급분화량개과정,정향급분화시함유상동기인고적세포불동기인표체적결과,기인표체수요일정적조건,포외기질적변화가인기세포형태학급기인표체방식적개변.목적:관찰골수기질세포재손상대서뇌조직제취액유도하,향신경원양세포분화적가능성.방법:취제5대전염록색형광단백적골수기질세포,분별용결혈재관주/정상대서뇌조직제취액진행유도분화배양,병설립공백대조.상차현미경하관찰세포형태변화,병행면역조직화학염색감정.결과여결론:원대배양적골수기질세포순화、확증후정균균일치적장사형,제3대세포균일표체CD44,CD106,불표체CD34.형광현미경하,록색형광단백전염후24 h가관찰도골수기질세포유형광표체,단강도초약;48 h후대다수세포발출명현록색형광.가입결혈재관주대서뇌조직제취액후,유도세포불부재형태상표현위신경원양특정,이차신경원특이성희순화매특이성항체정양성표체.여공백대조조비교,결혈재관주대서뇌조직제취액조화정상대서뇌조직제취액조골수기질세포분화솔균명현승고(P < 0.05),차전조승고폭도명현대우후조(P < 0.05).제시결혈재관주대서뇌조직제취액능장골수기질세포성공유도위신경원양세포.
BACKGROUND: Differentiation of bone marrow stromal cells (BMSCs) into neurons requires two processes: orientation and differentiation. Orientation and differentiation are results from different gene expression in cells with the same gene bank. Gene expression requires a certain condition. Changes in extracellular matrix can induce changes in cell morphology and gene expression manner. OBJECTIVE: To explore the possibility of BMSC differentiation into neuron-like cells under tissue extract from rat damaged brain. METHODS: The fifth passage of green fluorescent protein (GFP)-transfected BMSCs was induced to differentiate in brain tissue extract from ischemia/reperfusion rats or normal rats. A blank control was set. Cell morphology change was observed under the phase contrast microscope, and then evaluated using immunohistochemical staining. RESULTS AND CONCLUSION: Primarily cultured BMSCs were purified and amplified, and then showed even spindle shape. The third passage of BMSCs was positive for CD44 and CD106, but negative for CD34. Under the fluorescence microscope, BMSCs showed fluorescence expression, but the strength was weak 24 hours following GFP transfection. Numerous cells presented significant green fluorescence 48 hours later. Following adding brain tissue extract from ischemia/reperfusion rats. Induced cells presented neuron-like feature, but neuron specific enolase specific antibody presented positive expression. Compared with the blank control group, the differentiation rate of BMSCs was significantly increased in the ischemia/reperfusion group and normal group (P < 0.05). The increased range was significantly greater in the ischemia/reperfusion group than the normal group (P < 0.05). These results indicated that brain tissue extract from ischemia/reperfusion rats can successfully induce the differentiation of BMSCs into neuron-like cells.