Effects of basic fibroblast growth factor on the development of the stem cell properties of human dental pulp cells
Effects of basic fibroblast growth factor on the development of the stem cell properties of human dental pulp cells
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DOI：
10.1679/aohc.72.51
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发表时间：
2009-07-01
影响因子：
--
通讯作者：
Sato, Tetsuji
中科院分区：
文献类型：
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作者：
Morito, Akiyuki;Kida, Yujiro;Sato, Tetsuji
我们在胶原酶和分散酶处理人牙髓后分离粘附成纤维细胞。当人牙髓细胞（hDPC）培养在碱性成纤维细胞生长因子（bFGF）的存在下，hDPC的比例在S期是显着高于无bFGF的孵育相比。表达STRO-1作为干细胞群标志物的hDPC的比例在bFGF存在下增加约8倍，而不是在bFGF不存在下。我们证明了hDPC的特征和独特性，并表明，当与含有血清和bFGF的培养基一起培养时，它们具有高度增殖性，并且能够在体外分化为成骨细胞、软骨细胞和脂肪细胞。此外，在蛋白质和基因表达水平上证实了体外分化。将在bFGF存在下离体扩增的hDPC移植到免疫功能低下的小鼠中，显示骨、软骨和脂肪组织的形成。使用人特异性Alu探针在受体小鼠皮下组织中位于PLGA附近的骨组织中标记供体hDPC衍生的细胞。当用含有bFGF的无血清培养基培养时，hDPC强烈表达STRO-1免疫反应产物并持续自我更新，因此在分化潜能和增殖活性方面与用含有血清和bFGF的培养基培养的hDPC几乎相同。本结果表明，在bFGF的存在下培养的hDPC，无论牛血清的存在与否，都富含间充质干细胞或祖细胞，并且可用于治疗牙科疾病的基于细胞的疗法。
We isolated adherent fibroblastic cells after collagenase and dispase treatment of human dental pulp. When human dental pulp cells (hDPCs) were cultured in the presence of basic fibroblast growth factor (bFGF), the ratio of hDPCs in the S-phase was significantly higher in comparison with incubation without bFGF. The ratio of hDPCs expressing STRO-1 as a marker of stem cell populations increased approximately eightfold in the presence of bFGF as opposed to that in the absence of bFGF. We demonstrated the characterization and distinctiveness of the hDPCs and showed that, when cultured with the medium containing serum and bFGF, they were highly proliferative and capable of differentiating in vitro into osteoblasts, chondrocytes, and adipocytes. Furthermore, the in vitro differentiation was confirmed at both the protein and gene expression levels. Transplantation of hDPCs-expanded ex vivo in the presence of bFGF into immunocompromised mice-revealed the formation of bone, cartilage, and adipose tissue. The donor hDPC-derived cells were labeled in the bone tissues located near the PLGA in the subcutaneous tissues of recipient mice using a human-specific Alu probe. When cultured with a serum-free medium containing bFGF, the hDPCs strongly expressed STRO-1 immunoreactive products and sustained self-renewal, and thus were almost identical in differentiation potential and proliferation activity to hDPCs cultured with the medium containing serum and bFGF. The present results suggest that the hDPCs cultured in the presence of bFGF irrespective of the presence or absence of the bovine serum are rich in mesenchymal stem cells or progenitor cells and useful for cell-based therapies to treat dental diseases.