Cultivation of human embryonic stem cells without the embryoid body step enhances osteogenesis in vitro
Cultivation of human embryonic stem cells without the embryoid body step enhances osteogenesis in vitro
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DOI：
10.1634/stemcells.2005-0383
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发表时间：
2006-04-01
期刊：
影响因子：
5.2
通讯作者：
Langer, Robert S.
中科院分区：
文献类型：
--
作者：
Karp, Jeffrey M.;Ferreira, Lino S.;Langer, Robert S.
胚胎干细胞（ESC）的成骨培养主要来源于称为胚状体（EB）的三维细胞球体。另一种已经尝试并值得进一步关注的方法是通过将ESC集落立即分离成单细胞来避免EB。然而，这种方法尚未得到很好的表征，省略EB步骤的效果是未知的。在此，我们报告，培养人胚胎干细胞（hESC）没有EB阶段导致7倍以上的成骨细胞的数量和自发骨结节形成后10-12天。相比之下，当hESC分化为EB 5天，然后接种单细胞时，仅在地塞米松存在下4周后形成骨结节。此外，无论是否包含EB，骨基质形成，包括骨水泥线基质和矿化胶原，其显示具有与羟基磷灰石和人骨相似的钙磷比的磷灰石矿物（PO 4）。总之，这些结果表明，没有EB步骤的培养hESC可用于获得大量的功能性成骨细胞用于骨组织工程。
Osteogenic cultures of embryonic stem cells (ESCs) are predominately derived from three-dimensional cell spheroids called embryoid bodies (EBs). An alternative method that has been attempted and merits further attention avoids EBs through the immediate separation of ESC colonies into single cells. However, this method has not been well characterized and the effect of omitting the EB step is unknown. Herein, we report that culturing human embryonic stem cells (hESCs) without the EB stage leads to a sevenfold greater number of osteogenic cells and to spontaneous bone nodule formation after 10-12 days. In contrast, when hESCs were differentiated as EBs for 5 days followed by plating of single cells, bone nodules formed after 4 weeks only in the presence of dexamethasone. Furthermore, regardless of the inclusion of EBs, bone matrix formed, including cement line matrix and mineralized collagen, which displayed apatitic mineral (PO4) with calcium-to-phosphorous ratios similar to those of hydroxyapatite and human bone. Together these results demonstrate that culturing hESCs without an EB step can be used to derive large quantities of functional osteogenic cells for bone tissue engineering.