Isolation, characterization, and differentiation of stem cells derived from the rat amniotic membrane
Isolation, characterization, and differentiation of stem cells derived from the rat amniotic membrane
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DOI：
10.1111/j.1432-0436.2007.00194.x
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发表时间：
2008-02-01
期刊：
影响因子：
2.9
通讯作者：
Black, Ira B.
中科院分区：
文献类型：
--
作者：
Marcus, Akiva J.;Coyne, Thomas M.;Black, Ira B.
基于干细胞的疗法可以治疗多种疑难杂症。干细胞生物学的一个基本目标涉及表现出不同潜力、生长能力和治疗效用的不同群体的特征。我们报告了从大鼠羊膜组织外植体中分离出的干细胞群的特征。与间充质干细胞类似，这些羊膜源性干细胞 (ADSC) 表达表面标记物 CD29 和 CD90，但淋巴造血标记物 CD45 和 CD11b 呈阴性。 ADSC 以多分化状态存在于培养物中，表达神经外胚层（神经丝-M）、中胚层（纤连蛋白）和内胚层（α-1-抗胰蛋白酶）基因。为了评估可塑性，ADSC 接受了多种培养条件，旨在促进分化为神经外胚层、中胚层和内胚层细胞类型。在确定的神经诱导培养基中培养的 ADSC 呈现神经元形态并上调神经特异性基因。在不同条件下，ADSCs能够分化为假定的骨细胞和脂肪细胞，这分别通过矿化基质的沉积和积累的脂滴来表明。此外，在促进肝细胞分化的培养基中培养的 ADSC 上调肝脏特异性基因（白蛋白）和内化低密度脂蛋白（LDL），与肝细胞表型一致。为了确定观察到的可塑性是否反映了群体中真正干细胞的存在，我们从单个细胞中衍生出了个体克隆。克隆系再现了亲本 ADSC 培养物的表达模式，并且是多能的。 ADSC 已培养 20 代而没有失去其可塑性，表明其具有长期的自我更新能力。总之，我们的数据表明 ADSC 和衍生的克隆系能够长期自我更新和多分化，满足干细胞群的所有标准。
Stem-cell-based therapies may offer treatments for a variety of intractable diseases. A fundamental goal in stem-cell biology concerns the characterization of diverse populations that exhibit different potentials, growth capabilities, and therapeutic utilities. We report the characterization of a stem-cell population isolated from tissue explants of rat amniotic membrane. Similar to mesenchymal stem cells, these amnion-derived stem cells (ADSCs) express the surface markers CD29 and CD90, but were negative for the lymphohematopoietic markers CD45 and CD11b. ADSCs exist in culture in a multidifferentiated state, expressing neuroectodermal (neurofilament-M), mesodermal (fibronectin), and endodermal (alpha-1-antitrypsin) genes. To assess plasticity, ADSCs were subjected to a number of culture conditions intended to encourage differentiation into neuroectodermal, mesodermal, and endodermal cell types. ADSCs cultured in a defined neural induction media assumed neuronal morphologies and up-regulated neural-specific genes. Under different conditions, ADSCs were capable of differentiating into presumptive bone and fat cells, indicated by the deposition of mineralized matrix and accumulated lipid droplets, respectively. Moreover, ADSCs cultured in media that promotes liver cell differentiation up-regulated liver-specific genes (albumin) and internalized low-density lipoprotein (LDL), consistent with a hepatocyte phenotype. To determine whether this observed plasticity reflects the presence of true stem cells within the population, we have derived individual clones from single cells. Clonal lines recapitulate the expression pattern of parental ADSC cultures and are multipotent. ADSCs have been cultured for 20 passages without losing their plasticity, suggesting long-term self-renewal. In sum, our data suggest that ADSCs and derived clonal lines are capable of long-term self-renewal and multidifferentiation, fulfilling all the criteria of a stem-cell population.