CAJ | 학술논문

目的探讨异种骨对大鼠神经干细胞(NSCs)诱导分化的作用.方法从15 d胎龄大鼠的大脑皮质分离获得NSCs后,以神经球悬浮培养法,采用DMEM/F12为基础培养基,加入2%B27、20 μg/L EGF和20 μg/L bFGF等进行培养.传代3次后,采用巢蛋白染色,免疫荧光细胞化学法鉴定NSCs.然后,加入浓度分别为0.025、0.05、0.1、0.2 g/mL的异种骨浸提液与NSCs进行共培养.结果传代后的细胞表现出NSCs的特性.与0.2 g/mL异种骨浸提液共培养的NSCs微管相关蛋白、胶原纤维酸性蛋白和O4均呈阳性表达,即异种骨浸提液可显著诱导NSCs分化为神经元、星形胶质细胞和少突胶质细胞;其中异种骨浸提液浓度为0.1 g/mL时,诱导NSCs分化为少突胶质细胞的比例最高;浓度为0.05 g/mL时,分化为星形胶质细胞的比例最高.结论异种骨具有诱导NSCs分化为神经元及胶质细胞的作用,且浓度不同,诱导分化的细胞比例成分亦不同.临床应用中,可结合一定的生长因子,诱导其分化方向,从而有利于修复神经系统损伤.
목적 탐토이충골대대서신경간세포(NSCs)유도분화적작용.방법 종15 d태령대서적대뇌피질분리획득NSCs후,이신경구현부배양법,채용DMEM/F12위기출배양기,가입2%B27、20 μg/L EGF화20 μg/L bFGF등진행배양.전대3차후,채용소단백염색,면역형광세포화학법감정NSCs.연후,가입농도분별위0.025、0.05、0.1、0.2 g/mL적이충골침제액여NSCs진행공배양.결과 전대후적세포표현출NSCs적특성.여0.2 g/mL이충골침제액공배양적NSCs미관상관단백、효원섬유산성단백화O4균정양성표체,즉이충골침제액가현저유도NSCs분화위신경원、성형효질세포화소돌효질세포;기중이충골침제액농도위0.1 g/mL시,유도NSCs분화위소돌효질세포적비례최고;농도위0.05 g/mL시,분화위성형효질세포적비례최고.결론 이충골구유유도NSCs분화위신경원급효질세포적작용,차농도불동,유도분화적세포비례성분역불동.림상응용중,가결합일정적생장인자,유도기분화방향,종이유리우수복신경계통손상.
Objective To investigate the effect of xenogeneic bone on induced differentiation of neural stem cells (NSCs) .Methods NSCs were isolated from cerebral cortex of the frontal lobe in 15 d rat embryos and were cultured u-sing the neurosphere culture system.The basic culture medium was DMEM/F12,added with 2%B27,20μg/L EGF,20μg/L bFGF and others.After 3 passages,NSCs were stained by Nestin and identified by immunocytochemistry.They were then co-cultured with xenogeneic bone extracts at concentrations of 0.025,0.05,0.1,0.2 g/mL.Results Cells after passage exhibited characteristics of NSCs.Positive expression of microtubule-associated protein 2,glial fibrillary acidic protein and O4 were detected in NSCs co-cultured with 0.2 g/mL xenogeneic bone extracts.That is to say,xenogeneic bone extracts could induce NSCs differentiate into neurons,astrocytes and oligodendrocyte.When the concentration of xe-nogeneic bone extracts was 0.1 g/mL,the proportion of NSCs that differentiate into oligodendrocytes was the most.When the concentration was 0.05 g/mL,the proportion of NSCs that differentiate into astrocytes was the most.Conclusion Xe-nogeneic bone can induce NSCs to differentiate into neurons and glial cells.The proportion of differentiated cells was dif-ferent at various concentrations of xenogeneic bone.Therefore,in its clinical application,the xenogeneic bone combined with certain growth factors to induce NSC differentiation can help repair the damage of the nervous system.