Bradford Jolene A, Clarke Scott T
Life Technologies, Eugene, OR, USA.
Methods Mol Biol. 2011;698:367-85. doi: 10.1007/978-1-60761-999-4_27.
Adult human mesenchymal stem cells (hMSC) are rare fibroblast-like cells capable of differentiation into a variety of cell tissues which include bone, cartilage, muscle, ligament, tendon, and adipose. Normal adult bone marrow and adipose tissue are the most common sources of these cells. The International Society for Cellular Therapy (ISCT) has proposed a set of standards to define hMSC for laboratory investigations and preclinical studies: adherence to plastic in standard culture conditions; in vitro differentiation into osteoblasts, adipocytes, and chondroblasts; and specific surface antigen expression. Direct measurement of proliferation combined with simultaneous detection of the ISCT-consensus immunophenotypic profile provides data that is used to determine the differentiation status and health of the cells. Flow cytometry provides a powerful technology that is routinely used to simultaneously and rapidly measure multiple parameters in a single sample. This chapter describes a flow cytometric panel for the simultaneous detection of immunophenotypic profile, proliferative capacity, and DNA content measurement in hMSC. Because a relatively small number of cells are needed with this approach, measurements can be made with minimal impact on expansion potential. The ability to assess antigen expression and proliferative status enables the investigator to make informed decisions on expansion and harvesting.
成人人类间充质干细胞（hMSC）是一种罕见的成纤维细胞样细胞，能够分化为多种细胞组织，包括骨、软骨、肌肉、韧带、肌腱和脂肪组织。正常成人骨髓和脂肪组织是这些细胞最常见的来源。国际细胞治疗协会（ISCT）提出了一套标准，用于在实验室研究和临床前研究中定义hMSC：在标准培养条件下贴壁生长；体外分化为成骨细胞、脂肪细胞和成软骨细胞；以及特定表面抗原表达。直接测量增殖并同时检测ISCT共识免疫表型谱可提供用于确定细胞分化状态和健康状况的数据。流式细胞术提供了一种强大的技术，常用于在单个样本中同时快速测量多个参数。本章描述了一种流式细胞术检测方案，用于同时检测hMSC的免疫表型谱、增殖能力和DNA含量。由于这种方法所需的细胞数量相对较少，因此测量对扩增潜力的影响最小。评估抗原表达和增殖状态的能力使研究人员能够就细胞扩增和收获做出明智的决策。
