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Comparison of phenotypic markers and neural differentiation potential of multipotent adult progenitor cells and mesenchymal stem cells

机译:多能成体祖细胞和间充质干细胞的表型标记和神经分化潜能的比较

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摘要

AIM: To compare the phenotypic and neural differentiation potential of human bone marrow derived multipotent adult progenitor cells (MAPC) and mesenchymal stem cells (MSC).METHODS: Cultures of MAPC and MSC were established in parallel from same samples of human bone marrow (n = 5). Both stem cell types were evaluated for expression of pluripotency markers including Oct-4 and Nanog by immunocytochemistry and reverse-transcription polymerase chain reaction (RT-PCR) and expression of standard mesenchymal markers including CD14, CD34, CD44, CD45, CD73, CD90, CD105 and human leukocyte antigen (HLA)-ABC by flow cytometry. After treatment with neural induction medium both MAPC and MSC were evaluated for expression of neural proteins [neuronal filament-200 (NF-200) and glial fibrillar acidic protein (GFAP)] by immunocytochemistry and Western blotting and neural genes [NF-200, GFAP, Tau, microtubule-associated protein (MAP)-1B, MAP-2, neuron-specific enolase (NSE) and oligodendrocyte-1 (Olig-1)] by quantitative real-time-PCR.RESULTS: MAPC had small trigonal shaped while MSC had elongated spindle-shaped morphology. The MAPC expressed Oct-4 and Nanog both at gene and protein levels, whereas MSC were negative for these pluripotent markers. MAPC were negative for HLA-ABC while MSC had high expression of HLA-ABC. In addition, MAPC as compared to MSC had significantly lower expression of CD44 (36.56% ± 1.92% vs 98.23% ± 0.51%), CD73 (15.11% ± 2.24% vs 98.53% ± 2.22%) and CD105 (13.81% ± 3.82% vs 95.12% ± 5.65%) (P < 0.001, for all) MAPC cultures compared to MSC cultures treated with neural induction medium had significantly higher fold change expression of NF-200 (0.64), GFAP (0.52), Tau (0.59), MAP-2 (0.72), Olig-1 (0.18) and NSE (0.29) proteins (P < 0.01 for Olig-1 and P < 0.001 for rest) as well as higher fold change expression of genes of NF-200 (1.34), GFAP (1.12), Tau (1.08), MAP-1B (0.92), MAP-2 (1.14) and NSE (0.4) (P < 0.001 for all).CONCLUSION: MAPC can be differentially characterized from MSC as Oct-4 and Nanog positive stem cells with no expression of HLA-ABC and low expression of mesenchymal markers CD44, CD73 and CD105 and when compared to MSC they possess greater predilection for differentiation into neuro-ectodermal lineage.
机译:目的:比较人骨髓来源的多能成体祖细胞(MAPC)和间充质干细胞(MSC)的表型和神经分化潜能。方法:从相同的人骨髓样品中平行建立MAPC和MSC培养物(n = 5)。通过免疫细胞化学和逆转录聚合酶链反应(RT-PCR)评估了两种干细胞的多能性标记物(包括Oct-4和Nanog)的表达以及标准间质标记物(包括CD14,CD34,CD44,CD45,CD73,CD90,流式细胞仪检测CD105和人白细胞抗原(HLA)-ABC。在用神经诱导培养基处理后,通过免疫细胞化学和蛋白质印迹以及神经基因[NF-200,GFAP]评价了MAPC和MSC的神经蛋白[神经元细丝200(NF-200)和神经胶质纤维酸性蛋白(GFAP)]的表达。 ,Tau,微管相关蛋白(MAP)-1B,MAP-2,神经元特异性烯醇化酶(NSE)和少突胶质细胞1(Olig-1)]的定量实时荧光定量PCR。结果:MAPC具有小的三角形形状,而MSC具有细长的纺锤形形态。 MAPC在基因和蛋白质水平上都表达了Oct-4和Nanog,而MSC对这些多能标记物却是阴性的。 MAPC对HLA-ABC阴性,而MSC高表达HLA-ABC。此外,与MSC相比,MAPC的CD44(36.56%±1.92%对98.23%±0.51%),CD73(15.11%±2.24%对98.53%±2.22%)和CD105(分别为13.81%±3.82%)的表达明显降低。 vs 95.12%±5.65%)(对于所有P均<0.001)MAPC培养物与经神经诱导培养基处理的MSC培养物相比,其NF-200(0.64),GFAP(0.52),Tau(0.59), MAP-2(0.72),Olig-1(0.18)和NSE(0.29)蛋白(Olig-1 P <0.01,其余P <0.001)以及NF-200基因的更高倍数变化表达(1.34) ,GFAP(1.12),Tau(1.08),MAP-1B(0.92),MAP-2(1.14)和NSE(0.4)(所有P均<0.001)。结论:MAPC与MSC的区别在于Oct-4 Nanog阳性干细胞不表达HLA-ABC,间充质标记CD44,CD73和CD105的表达低。与MSC相比,它们更倾向于分化为神经外胚层谱系。

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