Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, 240 Skirkanich Hall, Philadelphia, Pennsylvania 19104, USA.
Biotechnol Bioeng. 2011 Jan;108(1):163-74. doi: 10.1002/bit.22925.
The use of high-throughput screening (HTS) techniques has long been employed by the pharmaceutical industry to increase discovery rates for new drugs that could be useful for disease treatment, yet this technology has only been minimally applied in other applications such as in tissue regeneration. In this work, an assay for the osteogenic differentiation of human mesenchymal stem cells (hMSCs) was developed and used to screen a library of small molecules for their potential as either promoters or inhibitors of osteogenesis, based on levels of alkaline phosphatase activity and cellular viability. From a library of 1,040 molecules, 36 promoters, and 20 inhibitors were identified as hits based on statistical criteria. Osteopromoters from this library were further investigated using standard culture techniques and a wider range of outcomes to verify that these compounds drive cellular differentiation. Several hits led to some improvement in the expression of alkaline phosphatase, osteogenic gene expression, and matrix mineralization by hMSCs when compared to the standard dexamethasone supplemented media and one molecule was investigated in combination with a recently identified biodegradable and osteoconductive polymer. This work illustrates the ability of HTS to more rapidly identify potential molecules to control stem cell differentiation.
高通量筛选 (HTS) 技术长期以来一直被制药行业用于提高新药的发现率，这些新药可能对疾病治疗有用，但这项技术在组织再生等其他应用中仅得到了最小程度的应用。在这项工作中，开发了一种用于人骨髓间充质干细胞 (hMSC) 成骨分化的测定法，并根据碱性磷酸酶活性和细胞活力水平，将小分子文库用于筛选潜在的成骨促进剂或抑制剂。根据统计标准，从 1040 种分子文库中鉴定出 36 种促进剂和 20 种抑制剂作为命中化合物。从该文库中选择的成骨促进剂，进一步使用标准培养技术和更广泛的结果进行了研究，以验证这些化合物可驱动细胞分化。与标准地塞米松补充培养基相比，一些化合物可使 hMSC 的碱性磷酸酶、成骨基因表达和基质矿化表达水平得到一定程度的提高，有一个化合物还与最近发现的一种可生物降解和骨传导聚合物进行了联合研究。这项工作说明了 HTS 能够更快速地识别潜在的分子来控制干细胞分化。
