Brown Shelley E, Tong Wilbur, Krebsbach Paul H
Biologic and Materials Sciences, School of Dentistry, University of Michigan, Ann Arbor, Mich. 48109-1078, USA.
Cells Tissues Organs. 2009;189(1-4):256-60. doi: 10.1159/000151746. Epub 2008 Aug 27.
Human embryonic stem cells (hESCs) hold promise for tissue regeneration therapies by providing a potentially unlimited source of cells capable of undergoing differentiation into specified cell types. Several preclinical studies and a few clinical studies use human bone marrow stromal cells (hBMSCs) to treat skeletal diseases and repair damaged tissue. However, hBMSCs have limited proliferation and differentiation capacity, suggesting that an alternate cell source is desirable, and hESCs may serve this purpose. Here we describe a protocol for the reproducible derivation of mesenchymal stem cells from hESCs (hES-MSCs). The hES-MSCs have a similar immunophenotype to hBMSCs, specifically they are CD73+, STRO-1+ and CD45-, and are karyotypically stable. The derived hES-MSCs are also capable of differentiating into osteoblasts and adipocytes. When the hES-MSCs were genetically modified with the lineage-specific Col2.3-GFP lentivirus and cultured in osteogenic medium, increased GFP expression was detected over time, indicating the hES-MSCs have the capacity to differentiate down the osteogenic lineage and had progressed toward a mature osteoblast phenotype.
人类胚胎干细胞（hESCs）通过提供一种潜在的无限细胞来源，有望用于组织再生治疗，这些细胞能够分化为特定的细胞类型。一些临床前研究和少数临床研究使用人类骨髓基质细胞（hBMSCs）来治疗骨骼疾病和修复受损组织。然而，hBMSCs的增殖和分化能力有限，这表明需要一种替代细胞来源，而hESCs可能符合这一需求。在此，我们描述了一种从hESCs（hES-MSCs）可重复衍生间充质干细胞的方案。hES-MSCs具有与hBMSCs相似的免疫表型，具体而言，它们是CD73+、STRO-1+和CD45-，并且核型稳定。所衍生的hES-MSCs也能够分化为成骨细胞和脂肪细胞。当用谱系特异性的Col2.3-GFP慢病毒对hES-MSCs进行基因改造并在成骨培养基中培养时，随着时间的推移检测到GFP表达增加，表明hES-MSCs具有沿着成骨谱系分化的能力，并已朝着成熟成骨细胞表型发展。
