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人胎儿视网膜祖细胞与脑源性神经干细胞体外分化的比较研究

[A comparison study for differentiation between human fetal retinal progenitor cells and brain neural stem cells in vitro].

作者信息

Kang Qian-yan, Liu Yong, Chen Xin-lin, Zhao Jian-jun, Zhang Peng-bo, Li Jie, Luo Yu, Qian Yi-hua, Song Tu-sheng

机构信息

Department of Ophthalmology, The First Affiliated Hospital, Xi'an Jiaotong University School of Medicine, Xi'an 710061, China.

出版信息

Zhonghua Yan Ke Za Zhi. 2006 Oct;42(10):901-7.

Abstract

OBJECTIVE

Investigating the potential of differentiation of human fetal retinal progenitor cells (hRPCs) and brain neural stem cells (hBNSCs) in vitro.

METHODS

hRPCs and hBNSCs were isolated from human fetuses (8-12 weeks of gestation) and cultured in serum-free DMEM/F12 culture medium with N2 supplement, epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) or culture medium with 10% fetal bovine serum (FBS) but without EGF and bFGF. Immunocytochemical and immunofluorescence studies were conducted for identification of neural stem cells, retinal progenitors or the subtypes of neurons, astrocytes, retinal ganglion cells and rod photoreceptors with the specific antibodies for Nestin, Pax6, Map2, GFAP, Thy-1 and Rhodopsin, respectively.

RESULTS

Both hRPCs and hBNSCs could proliferate and differentiate in DMEM/F12 + N2 with or without 10% FBS and expressed specific markers of immature neuroepithelial cells, retinal progenitors, mature neurons, astrocytes, retinal ganglion cells and rod photoreceptors. hBNSCs easily attached, spread out longer neurites and to form a network when cultured with serum contained medium. hRPCs were more difficult to attach and had only short dendrites.

CONCLUSIONS

Both hRPCs and hBNSCs can differentiate into retinal specific cell types in vitro. The adherent, migration and differential capacity of hRPCs and hBNSCs are different when these cells are induced by the serum-contained culture medium.

摘要

目的

研究人胎儿视网膜祖细胞(hRPCs)和脑神经元干细胞(hBNSCs)的体外分化潜能。

方法

从人胎儿(妊娠8 - 12周)中分离出hRPCs和hBNSCs,分别培养于添加N2、表皮生长因子(EGF)和碱性成纤维细胞生长因子(bFGF)的无血清DMEM/F12培养基中,或培养于含10%胎牛血清(FBS)但不含EGF和bFGF的培养基中。分别用针对巢蛋白(Nestin)、配对盒蛋白6(Pax6)、微管相关蛋白2(Map2)、胶质纤维酸性蛋白(GFAP)、甲状腺抗原1(Thy - 1)和视紫红质的特异性抗体进行免疫细胞化学和免疫荧光研究,以鉴定神经干细胞、视网膜祖细胞或神经元、星形胶质细胞、视网膜神经节细胞和视杆光感受器的亚型。

结果

hRPCs和hBNSCs在含或不含10% FBS的DMEM/F12 + N2培养基中均能增殖和分化,并表达未成熟神经上皮细胞、视网膜祖细胞、成熟神经元、星形胶质细胞、视网膜神经节细胞和视杆光感受器的特异性标志物。hBNSCs与含血清培养基一起培养时易于附着,长出更长的神经突并形成网络。hRPCs更难附着,只有短树突。

结论

hRPCs和hBNSCs在体外均可分化为视网膜特异性细胞类型。当用含血清培养基诱导时,hRPCs和hBNSCs的附着、迁移和分化能力有所不同。

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