Institute of Biomedical Engineering, Department of Engineering Science, University of Oxford, Oxford, United Kingdom.
Institute of Biomedical Engineering, Department of Engineering Science, University of Oxford, Oxford, United Kingdom; Department of Zoology, University of Oxford, Oxford, United Kingdom.
Biotechnol Adv. 2017 Jul;35(4):407-418. doi: 10.1016/j.biotechadv.2017.04.005. Epub 2017 Apr 24.
The plastic-adherent, fibroblast-like, clonogenic cells found in the human body now defined as multipotent "Mesenchymal Stromal Cells" (MSCs) hold immense potential for cell-based therapies. Recently, research and basic knowledge of these cells has fast-tracked, both from fundamental and translational perspectives. There have been important discoveries with respect to the available variety of tissue sources, the development of protocols for their easy isolation and in vitro expansion and for directed differentiation into various cell types. In addition, there has been discovery of novel abilities such as immune-modulation and further development of the use of biomaterials to aid isolation, expansion and differentiation together with improved delivery to the selected optimal tissue site. However, the molecular fingerprint of MSCs in these contexts remains imprecise and inadequate. Consequently, without this crucial knowledge it is difficult to achieve progress to determine with precision their practical developmental potentials. Detailed investigations on the global gene expression, or transcriptome, of MSCs could offer essential clues in this regard. In this article, we address the challenges associated with MSC transcriptome studies, the paradoxes observed in published experimental results and the need for careful transcriptomic analysis. We describe the exemplary applications with various transcriptome platforms that are used to address the variation in biomarkers and the identification of differentiation processes. The evolution and the potentials for adapting next-generation sequencing (NGS) technology in transcriptome analysis are discussed. Lastly, based on review of the existing understanding and published studies, we propose how NGS may be applied to promote further understanding of the biology of MSCs and their use in allied fields such as regenerative medicine.
目前在人体中发现的黏附塑料、成纤维样、克隆形成的细胞被定义为多能“间充质基质细胞”(MSCs),它们在基于细胞的治疗中具有巨大的潜力。最近,从基础和转化的角度来看,这些细胞的研究和基本知识都得到了快速发展。在组织来源的多样性、其易于分离和体外扩增的方案的开发以及向各种细胞类型的定向分化方面都有了重要的发现。此外,还发现了新的能力,如免疫调节,并进一步开发了生物材料的使用,以帮助分离、扩增和分化,同时改善向选定的最佳组织部位的输送。然而,在这些情况下,MSCs 的分子指纹仍然不够精确和不足。因此,没有这些关键的知识,就很难精确地确定它们的实际发展潜力。对 MSCs 的全局基因表达或转录组进行详细研究可能会为此提供重要线索。在本文中,我们将讨论与 MSC 转录组研究相关的挑战、在已发表的实验结果中观察到的悖论以及对转录组分析进行仔细分析的必要性。我们描述了使用各种转录组平台的典型应用,这些应用可用于解决生物标志物的变化和分化过程的鉴定问题。讨论了转录组分析中下一代测序(NGS)技术的发展和适应潜力。最后,基于对现有认识和已发表研究的综述,我们提出了 NGS 如何应用于促进对 MSCs 生物学的进一步理解及其在再生医学等相关领域的应用。