Induced pluripotent stem cell lines derived from human gingival fibroblasts and periodontal ligament fibroblasts
Induced pluripotent stem cell lines derived from human gingival fibroblasts and periodontal ligament fibroblasts
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DOI：
10.1111/j.1600-0765.2011.01358.x
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发表时间：
2011-08-01
影响因子：
3.5
通讯作者：
Bartold, P. M.
中科院分区：
文献类型：
--
作者：
Wada, N.;Wang, B.;Bartold, P. M.
背景和目的：人诱导多能干细胞（iPS）是由新生儿和成年人皮肤成纤维细胞通过重编程产生的，具有与人胚胎干细胞（hES）相似的特性。iPS细胞具有高的多能性和分化潜能，并可能是一个潜在的自体干细胞来源，为未来的再生therapy.Material和方法：iPS细胞系从人牙龈成纤维细胞，并为第一次，从牙周膜成纤维细胞，通过使用OCT 3/4，SOX 2，KLF4和c-MYC的逆转录病毒转导鸡尾酒重编程产生。通过胚胎干细胞标志物SSEA4、OCT 4、NANOG、GCTM-2、TG 30和TRA-1 - 60的表达研究iPS诱导。体外分化后，通过实时RT-PCR评估外胚层（SOX 1，PAX6）、中胚层[RUNX 1，T（Brachyury）]和内胚层（GATA 4，AFP）的分化标志物的基因表达。通过组织学评估植入小鼠睾丸后形成畸胎瘤的能力。结果：人牙龈成纤维细胞和牙周膜成纤维细胞来源的iPS细胞显示出与hES细胞相似的特征。两组iPS细胞均显示出与hES细胞相当的集落形态，并表达hES细胞相关的细胞表面抗原SSEA3、SSEA4、GCTM-2、TG30（CD9）和Tra-1 - 60，以及hES细胞标记基因OCT 4、NANOG和GDF 3。这些iPS细胞在体外表现出形成胚状体的分化潜能，并表达内胚层、外胚层和中胚层的基因。结论：成年人牙龈和牙周膜成纤维细胞可成功诱导产生iPS细胞。
Background and Objective: Human induced pluripotent stem (iPS) cells, which have similar properties to human embryonic stem (hES) cells, have been generated from neonatal and adult human dermal fibroblasts by reprogramming. iPS cells have high pluripotency and differentiation potential, and may be a potential autologous stem cell source for future regenerative therapy.Material and Methods: iPS cell lines from human gingival fibroblasts and, for the first time, from periodontal ligament fibroblasts, were generated by reprogramming using a retroviral transduction cocktail of OCT3/4, SOX2, KLF4 and c-MYC. iPS induction was investigated through expression of the embryonic stem cell markers SSEA4, OCT4, NANOG, GCTM-2, TG30 and TRA-1-60. Following in vitro differentiation, the expression of genes for differentiation markers for ectoderm (SOX1, PAX6), mesoderm [RUNX1, T(Brachyury)] and endoderm (GATA4, AFP) was assessed by real-time RT-PCR. The ability to form teratomas following implantation into mouse testes was assessed by histology.Results: Human gingival fibroblast-and periodontal ligament fibroblast-derived iPS cells showed similar characteristics to hES cells. Both sets of iPS cells displayed colony morphology comparable to that of hES cells and expressed the hES cell-associated cell-surface antigens, SSEA3, SSEA4, GCTM-2, TG30 (CD9) and Tra-1-60, and the hES cell marker genes, OCT4, NANOG and GDF3. These iPS cells showed differentiation potential to form embryoid bodies in vitro and expressed genes for endoderm, ectoderm and mesoderm. Teratoma formation following implantation into mouse testes was observed.Conclusion: These results demonstrate that iPS cells can be successfully generated from adult human gingival and periodontal ligament fibroblasts.