Spontaneous differentiation of mesenchymal stem cells obtained from fetal rat circulation
Spontaneous differentiation of mesenchymal stem cells obtained from fetal rat circulation
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DOI：
10.1016/j.bone.2004.05.006
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发表时间：
2004-10-01
期刊：
影响因子：
4.1
通讯作者：
Itoman, M
中科院分区：
文献类型：
--
作者：
Naruse, K;Urabe, K;Itoman, M
间充质干细胞（MSC）被认为是多潜能的，能够分化成多个谱系。我们试图表征来自胎儿循环血（FCBCs）的大鼠细胞，其显示成纤维细胞形态并分化为成骨细胞和软骨细胞谱系。值得注意的是，它们在塑料培养皿上在仅补充有10%胎牛血清（FBS）的培养基中分化成软骨细胞特异性表型，而不使用三维培养基质。骨髓来源的细胞在相同条件下不表达这种表型表达，这些细胞的特征通过逆转录聚合酶链反应、免疫组织化学和von Kossa染色以及免疫斑点印迹进行分析。在一个群体中，在分化的细胞中检测到11型和X型胶原蛋白的表达，其水平与在来自大鼠肋软骨的软骨细胞中观察到的水平相同。在另一个人群中，甲状旁腺激素受体，碱性磷酸酶，骨钙素的表达水平几乎等于长骨来源的成骨细胞中观察到的。培养3周后，广泛凝聚的细胞团，抗11型胶原抗体染色，可以区分从小，多层，冯Kossa阳性结节，抗骨钙素染色，但不与抗11型胶原抗体的组织学。此外，在甲基异丁基黄嘌呤、地塞米松、胰岛素和吲哚美辛的存在下，FCBCs分化为成脂细胞。这些细胞表达PPARgamma 2 mRNA，并通过油红-O染色检测到积累的脂质囊泡。我们的研究结果表明，FCBCs有潜力很容易分化成多个谱系，他们是不同的骨髓或循环血液来源的间充质干细胞从更成熟和成人在其自发分化的情况下，如转化生长因子-β（TGF-β）或地塞米松，或三维培养环境。(C)2004年爱思唯尔公司All rights reserved.
Mesenchymal stem cells (MSCs) are thought to be multipotential, capable of differentiating into multiple lineages. We attempted to characterize rat cells derived from fetal circulating blood (FCBCs) that displayed a fibroblastic morphology and differentiated into osteoblastic and chondrocytic lineages. Notably, they differentiated into a chondrocyte-specific phenotype on plastic culture dishes in medium supplemented only with 10% fetal bovine serum (FBS) without the use of a three-dimensional culture substrate. Bone marrow-derived cells did not convey such phenotypic expression under the same conditions.The characteristic features of these cells were analyzed by reverse transcription polymerase chain reaction, immunohistological and von Kossa staining, and by immuno-dot blotting. In one population, expression of collagen types 11 and X was detected in differentiated cells at the same levels as observed in chondrocytes derived from rat rib cartilage. In another population, parathyroid hormone receptor, alkaline phosphatase, and osteocalcin were also expressed at levels almost equal to those observed in long bone-derived osteoblasts. After 3 weeks in culture, extensively condensed cell masses, stained with anti-type 11 collagen antibody, could be distinguished histologically from small, multilayered, von Kossa-positive nodules, which stained with anti-osteocalcin, but not with anti-type 11 collagen antibody. In addition, the FCBCs differentiated into adipogenic cells in the presence of methyl-isobutyl xanthine, dexamethasone, insulin, and indomethacin. These cells expressed PPARgamma2 mRNA and accumulated lipid vesicles detectable by Oil red-O staining. Our findings suggest that FCBCs have the potential to readily differentiate into multiple lineages and that they are distinct from mesenchymal stem cells derived from bone marrow or circulating blood from more mature and adults in their spontaneous differentiation in the absence of specific factors such as transforming growth factor-beta (TGF-beta) or dexamethasone, or a three-dimensional culture environment. (C) 2004 Elsevier Inc. All rights reserved.