Generation of cleidocranial dysplasia-specific human induced pluripotent stem cells in completely serum-, feeder-, and integration-free culture.
Generation of cleidocranial dysplasia-specific human induced pluripotent stem cells in completely serum-, feeder-, and integration-free culture.
复制标题
DOI：
10.1007/s11626-015-9968-x
复制
发表时间：
2016-02
期刊：
影响因子：
--
通讯作者：
Okamoto T
中科院分区：
文献类型：
--
作者：
Yamasaki S;Hamada A;Akagi E;Nakatao H;Ohtaka M;Nishimura K;Nakanishi M;Toratani S;Okamoto T
人类多能干细胞具有巨大的应用和科学潜力。为了提高对自我更新和分化的理解，我们先前报道了一种确定的无血清培养基hESF 9可以使用逆转录病毒载体在无血清和无饲养层的培养条件下产生和维持人诱导多能干细胞（iPSC）。为了避免与逆转录病毒整合相关的不可预测的副作用，我们在这里报告了用表达四个关键重编程基因的非整合复制缺陷型和持续性仙台病毒（SeVdp）载体从牙髓细胞成功产生hiPSC。我们发现hESF 9培养基与纤连蛋白的组合对于产生和维持具有SeVdp的hiPSC（KOSM）是有效的。使用该系统，多能和自我更新的hiPSC可以容易且稳定地产生和繁殖。利用该系统，我们成功地从由侏儒相关蛋白2（RUNX 2）的杂合种系突变引起的锁骨颅骨发育不良（CCD）中产生了hiPSCs，RUNX 2在成骨细胞的分化和软骨细胞的成熟中具有重要作用。这是建立CCD特异性iPSCs的第一份报告。CCD-iPSC畸胎瘤中的软骨显示异常。这些CCD-iPSCs将有助于阐明其分子机制和开发医学应用。此外，它带来了新的病理生理作用的RUNX 2在人类软骨细胞和骨细胞的分化。本文的在线版本（doi：10.1007/s11626-015-9968-x）包含补充材料，可供授权用户使用。
Human pluripotent stem cells hold great promise for their practical and scientific potentials. To improve understanding of self-renewal and differentiation, we previously reported a defined serum-free medium hESF9 could generate and maintain human induced pluripotent stem cells (iPSCs) in serum- and feeder-free culture conditions using retroviral vectors. To avoid the unpredictable side effects associated with retrovirus integration, we report here the successful generation of hiPSCs from dental pulp cells with a non-integrating replication-defective and persistent Sendai virus (SeVdp) vector expressing four key reprogramming genes. We found that hESF9 medium in combination with fibronectin are effective for generating and maintaining hiPSCs with SeVdp (KOSM). Using this system, pluripotent and self-renewing hiPSCs could be easily and stably generated and propagated. With this system, we successfully generated hiPSCs from cleidocranial dysplasia (CCD) caused by a heterozygous germ-line mutation of runt-related protein2 (RUNX2), which has an important role in the differentiation of osteoblasts and maturation of chondrocytes. This is the first report of the establishment of CCD-specific iPSCs. The cartilage in the teratomas of CCD-iPSCs showed abnormalities. These CCD-iPSCs would be beneficial to clarify the molecular mechanism and for development of medical applications. Moreover, it brings new pathophysiological role of RUNX2 in the differentiation of the human chondrocytes and osteocytes. The online version of this article (doi:10.1007/s11626-015-9968-x) contains supplementary material, which is available to authorized users.