CAJ | 학술논문

目的探讨新生大鼠脊髓来源神经干细胞(NSCs)的分离培养及在体外一定条件下向周围神经雪旺氏细胞分化的可行性. 方法分离新生大鼠的脊髓组织,在含有B27(终浓度1%)、碱性成纤维细胞生长因子(bFGF)和表皮生长因子(EGF)(终浓度均为20 μg/L)培养基中分离培养出NSCs.用复合诱导因子(10%FBS+5 μmol/L血小板凝集抑制剂+10 ng/mL bFGF+5 ng/mE血小板源性生长因子)在体外诱导NSCs分化为雪旺氏细胞.免疫荧光细胞化学方法[一抗为p75、S-100、神经胶质纤维酸性蛋白(GFAP)]鉴定体外诱导分化结果.结果培养的新生大鼠脊髓组织细胞nestin染色表达阳性;分离培养的大鼠脊髓来源NSCs经诱导分化后形态类似雪旺氏细胞,免疫荧光细胞化学方法显示诱导后细胞表达雪旺氏细胞的表面标志,GFAP、S-100和P75表达阳性.结论新生大鼠脊髓来源NSCs可以在体外诱导分化为雪旺氏细胞.
목적 탐토신생대서척수래원신경간세포(NSCs)적분리배양급재체외일정조건하향주위신경설왕씨세포분화적가행성. 방법 분리신생대서적척수조직,재함유B27(종농도1%)、감성성섬유세포생장인자(bFGF)화표피생장인자(EGF)(종농도균위20 μg/L)배양기중분리배양출NSCs.용복합유도인자(10%FBS+5 μmol/L혈소판응집억제제+10 ng/mL bFGF+5 ng/mE혈소판원성생장인자)재체외유도NSCs분화위설왕씨세포.면역형광세포화학방법[일항위p75、S-100、신경효질섬유산성단백(GFAP)]감정체외유도분화결과.결과 배양적신생대서척수조직세포nestin염색표체양성;분리배양적대서척수래원NSCs경유도분화후형태유사설왕씨세포,면역형광세포화학방법현시유도후세포표체설왕씨세포적표면표지,GFAP、S-100화P75표체양성.결론 신생대서척수래원NSCs가이재체외유도분화위설왕씨세포.
Objective To study whether peripheral nerve Schwann cells can be derived from neural stem cells obtained from neonatal rat spinal cord under certain condition in vitro. Methods Cells of neonatal rat spinal cord were isolated and maintained in conditioned medium that was supplemented with B27(final concentration of 1%),recombinant hunman basic-fibroblast growth factor (bFGF)and epidermal growth factor(EGF,final concentration of 20 μg/L)to induce the neural stem cells.Schwann cells were induced subsequently by compound induce factors(5 mL forskolin+10 ng/mL basic-FGF+5 ng/mL PDGF-AA+10%FBS).Immunohistochemical staining(antibody:p75,S-100 and GFAP) was performed to indentify the induction of Schwann cells. Results Cells isolated from neonatal rat spinal cord showed positive expression of nestin;neural stem cells induced from these cells can subsequently differentiate into morphologically Schwann cell-like cells in the supplements of compound factors.Positive expressions of Schwann cells markers(GFAP,S-100 and P75)were detected after immunofluorescence cytochemical staining of these cells.Conclusion Neural stem cells obtained from neonatal rat spinal cord can be differentiated into Schwana cells under the certain conditioned invitro culture.