Isolation and characterization of bone marrow-derived mesenchymal progenitor cells with myogenic and neuronal properties
Isolation and characterization of bone marrow-derived mesenchymal progenitor cells with myogenic and neuronal properties
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DOI：
10.1016/j.yexcr.2006.12.017
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发表时间：
2007-03-10
影响因子：
3.7
通讯作者：
Nakahata, Tatsutoshi
中科院分区：
文献类型：
--
作者：
Shiota, Mitsutaka;Heike, Toshio;Nakahata, Tatsutoshi
球体形成已被用作从各种组织中分离多能干/祖细胞的方法。然而，很少有关于骨髓衍生球的研究发表并评估其多潜能性。在这项研究中，使用三步法分离多能骨髓细胞群。首先，在去除造血细胞后，在塑料培养皿中培养小鼠骨髓来源的贴壁细胞，直到小细胞逐渐出现并增殖。然后在非粘附条件下培养细胞，并形成对神经前体标记物巢蛋白呈免疫阳性的球体。RT-PCR分析还显示，除了PPAR γ、osf 2、SOX 9和myoD外，球体对巢蛋白呈阳性，它们分别是脂肪细胞、成骨细胞、软骨细胞和骨骼成髓细胞谱系的前体标志物。最后，将球体解离成单细胞并在贴壁培养物中扩增。在适当的诱导条件下，球衍生的细胞获得的表型特性在体外的神经元，骨骼肌成肌细胞，跳动的心肌细胞，以及脂肪细胞，成骨细胞和软骨细胞。接下来，将球体衍生的细胞移植到小鼠心肌梗死模型中。一个月后，他们已经成为移植的心肌细胞，心导管检查显示显着的功能改善。因此，球源性细胞代表了一种新的方法，以提高小鼠骨髓的多分化潜能。(c)2007爱思唯尔公司All rights reserved.
Sphere formation has been utilized as a way to isolate multipotent stem/progenitor cells from various tissues. However, very few studies on bone marrow-derived spheres have been published and assessed their multipotentiality. In this study, multipotent marrow cell populations were isolated using a three-step method. First, after elimination of hematopoietic cells, murine marrow-derived adherent cells were cultured in plastic dishes until small cells gradually appeared and multiplied. Cells were then cultured under non-adherent conditions and formed spheres that were immunopositive for a neural precursor marker, nestin. RT-PCR analysis also revealed that the spheres were positive for nestin in addition to PPAR gamma, osf2, SOX9, and myoD, which are markers of precursors of adipocytic, osteoblastic, chondrocytic, and skeletal myeloblastic lineages, respectively. Finally, spheres were dissociated into single cells and expanded in adherent cultures. Under appropriate induction conditions, the sphere-derived cells acquired the phenotypic properties in vitro of neurons, skeletal myoblasts, and beating cardiomyocytes, as well as adipocytes, osteoblasts, and chondrocytes. Next, spherederived cells were transplanted into murine myocardial infarction models. One month later, they had become engrafted as cardiornyocytes, and cardiac catheterization showed significant functional improvements. Thus, sphere-derived cells represent a new approach to enhance the multi-differentiation potential of murine bone marrow. (c) 2007 Elsevier Inc. All rights reserved.