Insulin-Like Growth Factor 2 Promotes Osteogenic Cell Differentiation in the Parthenogenetic Murine Embryonic Stem Cells
Insulin-Like Growth Factor 2 Promotes Osteogenic Cell Differentiation in the Parthenogenetic Murine Embryonic Stem Cells
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DOI：
10.1089/ten.tea.2011.0074
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发表时间：
2012-02-01
影响因子：
4.1
通讯作者：
Roh, Sangho
中科院分区：
文献类型：
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作者：
Kang, Hoin;Sung, Jihye;Roh, Sangho
胚胎干细胞（ESC）是多能的，可以分化成所有的体细胞类型。ESC是硬组织再生和骨骼组织修复的替代解决方案，用于使用再生策略治疗骨疾病和缺损。单性胚胎干细胞（PESCs）可能是一个有用的替代干细胞来源的组织修复和再生。这种细胞类型在全期发育中的缺陷使研究人员能够避免与ESC研究相关的伦理问题。此外，在女性患者中，如果PESC来自卵母细胞，那么它们将具有该患者的遗传信息。在这里，我们提出的数据表明，成骨分化的PESCs可以促进胰岛素样生长因子2（IGF 2）。将PESC铺在具有补充有或没有IGF 2的ESC培养基的皮氏培养皿上，随后培养细胞1周。PESC形成漂浮的聚集体，称为胚状体（EB）。通过将EB在含有血清、抗坏血酸、β-甘油磷酸盐和视黄酸的培养基中孵育20天（含或不含IGF 2）诱导成骨谱系。通过实时聚合酶链反应分析特定成骨细胞标志物的基因表达，如骨钙素、骨桥蛋白、骨连接蛋白、骨唾液蛋白、I型胶原、碱性磷酸酶和Runx 2（Cbfa-I）。IGF 2处理的PESC衍生物中骨钙素、骨桥蛋白、骨连接蛋白和碱性磷酸酶的表达水平是IGF 2未处理的PESC衍生物的两倍。还使用临界大小的颅骨缺损小鼠模型进行体内实验。细胞移植后10周，IGF 2处理的PESC移植组比IGF 2未处理的PESC移植组观察到更多的骨组织再生。我们的体外和体内数据都表明，IGF 2诱导PESC的成骨分化。添加IGF 2可以重新激活PESC中的印记基因，这些基因仅在父亲基因组中表达，并且通常在PESC中沉默。我们的研究结果为骨骼组织修复机制和干细胞中的印迹机制提供了深入的见解。
Embryonic stem cells (ESCs) are pluripotent and can differentiate into all somatic cell types. ESCs are an alternative solution to hard tissue regeneration and skeletal tissue repair to treat bone diseases and defects using regenerative strategies. Parthenogenetic ESCs (PESCs) may be a useful alternative stem cell source for tissue repair and regeneration. The defects in full-term development of this cell type enable researchers to avoid the ethical concerns related to ESC research. Moreover, in female patients, if the PESCs are derived from oocytes, then they will have that patient's genetic information. Here, we present data demonstrating that osteogenic differentiation of PESCs can be promoted by insulin-like growth factor 2 (IGF2). PESCs were plated onto Petri dishes with ESC culture medium supplemented with or without IGF2, followed by culturing of the cells for 1 week. PESCs formed floating aggregates called embryoid bodies (EBs). An osteogenic lineage was induced from the EBs by incubating them in medium containing serum, ascorbic acid, b-glycerophosphate, and retionic acid, with or without IGF2, for 20 days. Gene expression of specific osteoblastic markers such as osteocalcin, osteopontin, osteonectin, bone sialoprotein, collagen type-I, alkaline phosphatase, and Runx2 (Cbfa-I) was analyzed by real-time polymerase chain reaction. The expression level of osteocalcin, osteopontin, osteonectin, and alkaline phosphatase was twofold higher in IGF2-treated PESC derivatives than IGF2-naive PESC derivatives. In vivo experiments were also performed using a critical-sized calvarial defect mouse model. Ten weeks after cell transplantation, more bone tissue regeneration was observed in the IGF2-treated PESC transplantation group than in IGF2-naive PESC transplantation group. Both our in vitro and in vivo data indicate that IGF2 induces osteogenic differentiation of PESCs. Addition of IGF2 may reactivate imprinting genes in PESCs that are only expressed in the paternal genome and are normally silent in PESCs. Our findings provide insights into the mechanisms of skeletal tissue repair and the imprinting mechanisms active in stem cells.