Reprogramming of mesenchymal stem cells derived from iPSCs seeded on biofunctionalized calcium phosphate scaffold for bone engineering.
Reprogramming of mesenchymal stem cells derived from iPSCs seeded on biofunctionalized calcium phosphate scaffold for bone engineering.
复制标题
对接种在生物功能化磷酸钙支架上的 iPSC 衍生的间充质干细胞进行重编程，用于骨工程。
DOI：
10.1016/j.biomaterials.2013.07.029
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发表时间：
2013-10
期刊：
影响因子：
14
通讯作者：
Xu, Hockin H. K.
中科院分区：
文献类型：
--
作者：
Liu, Jun;Chen, Wenchuan;Zhao, Zhihe;Xu, Hockin H. K.
关键词：
人诱导多能干细胞衍生的间充质干细胞（iPSC-MSC）是患者特异性干细胞的一个有前途的选择，具有卓越的细胞扩增能力。目前尚未见关于iPSC-MSC的骨形态发生蛋白2（BMP2）基因修饰用于骨组织工程的报道。本研究的目的是：（1）对 iPSC-MSC 进行基因改造以用于 BMP2 传递； (2) 将 BMP2 基因修饰的 iPSC-MSC 接种在固定有 RGD 的磷酸钙水泥 (CPC) 上，用于骨组织工程。 iPSC-MSCs 用绿色荧光蛋白 (GFP-iPSC-MSCs) 或 BMP2 慢病毒 (BMP2-iPSC-MSCs) 感染。检测到高水平的 GFP 表达，超过 68% 的 GFP-iPSC-MSC 呈 GFP 阳性。在定量 RT-PCR 和 ELISA 测定中，BMP2-iPSC-MSC 表达的 BMP2 水平高于 iPSC-MSC（p < 0.05）。与 iPSC-MSC 相比，BMP2-iPSC-MSC 不会影响生长动力学和细胞周期阶段。在成骨培养基中14天后，BMP2-iPSC-MSCs的ALP活性是iPSC-MSCs的1.8倍(p < 0.05)，表明iPSC-MSCs的BMP2基因转导增强了成骨分化。 BMP2-iPSC-MSC 接种在用 RGD 生物功能化的 CPC 支架上 (RGD-CPC)。 BMP2-iPSC-MSC 在 RGD-CPC 上附着良好。 14 d时，BMP2-iPSC-MSCs的COL1A1表达量是iPSC-MSCs的1.9倍。 BMP2-iPSC-MSCs的OC表达量是iPSC-MSCs的2.3倍。 21天时BMP2-iPSC-MSCs的骨基质矿化是iPSC-MSCs的1.8倍。总之，接种在 CPC 上的 iPSC-MSC 适用于骨组织工程。 RGD-CPC 上 BMP2 基因修饰的 iPSC-MSC 经历成骨分化，iPSC-MSC 中 BMP2 的过表达增强了 RGD-CPC 上的分化和骨矿物质生成。 BMP2-iPSC-MSC 接种在 RGD-CPC 支架上有望增强骨再生功效。
Human induced pluripotent stem cell-derived mesenchymal stem cells (iPSC-MSCs) are a promising choice of patient-specific stem cells with superior capability of cell expansion. There has been no report on bone morphogenic protein 2 (BMP2) gene modification of iPSC-MSCs for bone tissue engineering. The objectives of this study were to: (1) genetically modify iPSC-MSCs for BMP2 delivery; and (2) to seed BMP2 gene-modified iPSC-MSCs on calcium phosphate cement (CPC) immobilized with RGD for bone tissue engineering. iPSC-MSCs were infected with green fluorescence protein (GFP-iPSC-MSCs), or BMP2 lentivirus (BMP2-iPSC-MSCs). High levels of GFP expression were detected and more than 68% of GFP-iPSC-MSCs were GFP positive. BMP2-iPSC-MSCs expressed higher BMP2 levels than iPSC-MSCs in quantitative RT-PCR and ELISA assays (p < 0.05). BMP2-iPSC-MSCs did not compromise growth kinetics and cell cycle stages compared to iPSC-MSCs. After 14 d in osteogenic medium, ALP activity of BMP2-iPSC-MSCs was 1.8 times that of iPSC-MSCs (p < 0.05), indicating that BMP2 gene transduction of iPSC-MSCs enhanced osteogenic differentiation. BMP2-iPSC-MSCs were seeded on CPC scaffold biofunctionalized with RGD (RGD-CPC). BMP2-iPSC-MSCs attached well on RGD-CPC. At 14 d, COL1A1 expression of BMP2-iPSC-MSCs was 1.9 times that of iPSC-MSCs. OC expression of BMP2-iPSC-MSCs was 2.3 times that of iPSC-MSCs. Bone matrix mineralization by BMP2-iPSC-MSCs was was 1.8 times that of iPSC-MSCs at 21 d. In conclusion, iPSC-MSCs seeded on CPC were suitable for bone tissue engineering. BMP2 gene-modified iPSC-MSCs on RGD-CPC underwent osteogenic differentiation, and the overexpression of BMP2 in iPSC-MSCs enhanced differentiation and bone mineral production on RGD-CPC. BMP2-iPSC-MSC seeding on RGD-CPC scaffold is promising to enhance bone regeneration efficacy.
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DOI：
10.1002/jbm.1074
发表时间：
2002-02-01
期刊：
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH
影响因子：
--
作者：
Barralet, JE;Gaunt, T;Knowles, JC
通讯作者：
Knowles, JC
影响因子：
--
作者：
Johnson, Peter C.;Mikos, Antonios G.;Jansen, John A.
通讯作者：
Jansen, John A.
影响因子：
7.6
作者：
Alsberg, E;Kong, HJ;Mooney, DJ
通讯作者：
Mooney, DJ
影响因子：
158.5
作者：
BLAU, HM;SPRINGER, ML
通讯作者：
SPRINGER, ML
影响因子：
4.1
作者：
Hsu, W. K.;Sugiyama, O.;Lieberman, J. R.
通讯作者：
Lieberman, J. R.