Transplantation of Osteogenically Differentiated Mouse iPS Cells for Bone Repair
Transplantation of Osteogenically Differentiated Mouse iPS Cells for Bone Repair
复制标题
DOI：
10.3727/096368911x605529
复制
发表时间：
2012-01-01
影响因子：
3.3
通讯作者：
Ozaki, Toshifumi
中科院分区：
文献类型：
--
作者：
Hayashi, Takahiro;Misawa, Haruo;Ozaki, Toshifumi
诱导多能干细胞（iPS）是一种未分化的细胞，可以从已分化的细胞中获得，具有多能分化成肌肉骨骼系统、心肌、血管内皮细胞、神经元和肝细胞的潜能。因此，我们将小鼠iPS细胞在含有15%胎牛血清、10(-7)M地塞米松、10 mM β -甘油磷酸酯和50 μ g/ml抗坏血酸的DMEM中培养3周，以诱导骨分化，并采用RT-PCR技术以时间依赖性的方式研究骨分化标志物Runx2和骨钙素的表达。骨钙素是骨形成过程中的骨分化标志物，在第三周表达最高。用茜素红S染色观察钙结节沉积。将培养成骨分化的iPS细胞移植到严重联合免疫缺陷（SCID）小鼠体内，观察其4周后的成骨潜能。当骨分化诱导的iPS细胞移植到SCID小鼠体内时，在软x线图像和组织标本中证实骨形成。然而，20%的移植模型证实形成畸胎瘤。当小鼠iPS细胞在移植前接受2gy辐射处理时，畸胎瘤的形成受到抑制。当以同样方式处理的小鼠iPS细胞被移植到大鼠体内时，证实了骨形成，但未观察到畸胎瘤形成。认为移植前照射是抑制畸胎瘤形成的有效途径。
Induced pluripotent stem (iPS) cells are a type of undifferentiated cell that can be obtained from differentiated cells and have the pluripotent potential to differentiate into the musculoskeletal system, the myocardium, vascular endothelial cells, neurons, and hepatocytes. We therefore cultured mouse iPS cells in a DMEM containing 15% FBS, 10(-7) M dexamethasone, 10 mM beta-glycerophosphate, and 50 mu g/ml ascorbic acid for 3 weeks, in order to induce bone differentiation, and studied the expression of the bone differentiation markers Runx2 and osteocalcin using RT-PCR in a time-dependent manner. Osteocalcin, a bone differentiation marker in bone formation, exhibited the highest expression in the third week. In addition, the deposition of calcium nodules was observed using Alizarin red S staining. iPS cells cultured for bone differentiation were transplanted into severe combined immunodeficiency (SCID) mice, and the osteogenic potential exhibited after 4 weeks was studied. When bone differentiation-induced iPS cells were transplanted into SCID mice, bone formation was confirmed in soft X-ray images and tissue specimens. However, teratoma formation was confirmed in 20% of the transplanted models. When mouse iPS cells were treated with irradiation of 2 Gray (Gy) prior to transplantation, teratoma formation was inhibited. When mouse iPS cells treated in a likewise manner were xenotransplanted into rats, bone formation was confirmed but teratoma formation was not observed. It is believed that irradiation before transplantation is an effective way to inhibit teratoma formation.