山东医药
山東醫藥
산동의약
SHANDONG MEDICAL JOURNAL
2014年
47期
17-20
,共4页
生长因子%脂肪干细胞%神经细胞
生長因子%脂肪榦細胞%神經細胞
생장인자%지방간세포%신경세포
growth factor%adipose stem cells%nerve cells
目的：观察生长因子分步诱导法在大鼠脂肪干细胞（ADSCs）向神经细胞分化中的应用效果。方法取SD大鼠腹股沟及腹膜后脂肪，体外分离培养ADSCs，取消化、离心的第3代ADSCs，首先以DMEM／F12＋2％B27＋20 ng／mL EGF＋20 ng／mL bFGF的神经干细胞培养基诱导7 d，后以DMEM／F12＋1μmol／L RA神经诱导培养基诱导7 d。免疫荧光法鉴定分化后神经细胞特异标志物中神经细胞特异标志物巢蛋白（ Nestin ）、微管相关蛋白2（MAP2）、神经元核抗原抗体（NeuN）、神经胶质纤维酸性蛋白（GFAP）。结果从大鼠脂肪组织中成功提取AD-SCs，其能够连续传代且稳定增殖；流式细胞检测显示CD34表达率为0．36％，CD73为99．17％，CD90为98．61％，CD105为96．20％。经生长因子诱导7 d后细胞聚集成球，Nestin高表达；14 d后MAP2、NeuN高表达，而GFAP低表达。结论 ADSCs能够稳定增殖传代，生长因子分步诱导可高效诱导ADSCs定向分化为神经细胞。
目的：觀察生長因子分步誘導法在大鼠脂肪榦細胞（ADSCs）嚮神經細胞分化中的應用效果。方法取SD大鼠腹股溝及腹膜後脂肪，體外分離培養ADSCs，取消化、離心的第3代ADSCs，首先以DMEM／F12＋2％B27＋20 ng／mL EGF＋20 ng／mL bFGF的神經榦細胞培養基誘導7 d，後以DMEM／F12＋1μmol／L RA神經誘導培養基誘導7 d。免疫熒光法鑒定分化後神經細胞特異標誌物中神經細胞特異標誌物巢蛋白（ Nestin ）、微管相關蛋白2（MAP2）、神經元覈抗原抗體（NeuN）、神經膠質纖維痠性蛋白（GFAP）。結果從大鼠脂肪組織中成功提取AD-SCs，其能夠連續傳代且穩定增殖；流式細胞檢測顯示CD34錶達率為0．36％，CD73為99．17％，CD90為98．61％，CD105為96．20％。經生長因子誘導7 d後細胞聚集成毬，Nestin高錶達；14 d後MAP2、NeuN高錶達，而GFAP低錶達。結論 ADSCs能夠穩定增殖傳代，生長因子分步誘導可高效誘導ADSCs定嚮分化為神經細胞。
목적：관찰생장인자분보유도법재대서지방간세포（ADSCs）향신경세포분화중적응용효과。방법취SD대서복고구급복막후지방，체외분리배양ADSCs，취소화、리심적제3대ADSCs，수선이DMEM／F12＋2％B27＋20 ng／mL EGF＋20 ng／mL bFGF적신경간세포배양기유도7 d，후이DMEM／F12＋1μmol／L RA신경유도배양기유도7 d。면역형광법감정분화후신경세포특이표지물중신경세포특이표지물소단백（ Nestin ）、미관상관단백2（MAP2）、신경원핵항원항체（NeuN）、신경효질섬유산성단백（GFAP）。결과종대서지방조직중성공제취AD-SCs，기능구련속전대차은정증식；류식세포검측현시CD34표체솔위0．36％，CD73위99．17％，CD90위98．61％，CD105위96．20％。경생장인자유도7 d후세포취집성구，Nestin고표체；14 d후MAP2、NeuN고표체，이GFAP저표체。결론 ADSCs능구은정증식전대，생장인자분보유도가고효유도ADSCs정향분화위신경세포。
Objective To investigate the effect of rat adipose tissue-derived cells ( ADSCs) into neural cells with a growth factor two-step induction method .Methods ADSCs were isolated from the fat pads in inguinal and retroperitoneal of Sprague Dawley rats , and the third generation cells were cultured .First, the cells were induced with DMEM/F12+2%B27+20 ng/mL EGF+20 ng/mL bFGF for 7 days, and then DMEM/F12+1μmol/L RA for another 7 days.Immunocyto-chemistry was taked to identify the expression of the neuron specific markers Nestin , MAP2, NeuN and GFAP.Results ADSCs were reaped from rat fat tissue , and could passage in a considerable large amount .ADSCs could strongly express CD73(99.17%), CD90(98.61%), CD105(96.20%)and lowy expressed CD34(0.36%).The ADSCs induced for 7 days aggregated as floating cell spheres and positive for Nestin , then strongly express MAP2 and NeuN, and lowy expressed GFAP after 14 days.Conclusion The growth factor two-step induction method can efficiently induce ADSCs to differenti-ate into nerve cells .
