San Diego Heart Research Institute, San Diego State University, San Diego, USA.
Frederick National Laboratory for Cancer Science, Washington, USA.
Sci Rep. 2018 Aug 13;8(1):12060. doi: 10.1038/s41598-018-30551-1.
Regenerative therapeutic approaches for myocardial diseases often involve delivery of stem cells expanded ex vivo. Prior studies indicate that cell culture conditions affect functional and phenotypic characteristics, but relationship(s) of cultured cells derived from freshly isolated populations and the heterogeneity of the cultured population remain poorly defined. Functional and phenotypic characteristics of ex vivo expanded cells will determine outcomes of interventional treatment for disease, necessitating characterization of the impact that ex vivo expansion has upon isolated stem cell populations. Single-cell RNA-Seq profiling (scRNA-Seq) was performed to determine consequences of culture expansion upon adult cardiac progenitor cells (CPCs) as well as relationships with other cell populations. Bioinformatic analyses demonstrate that identity marker genes expressed in freshly isolated cells become undetectable in cultured CPCs while low level expression emerges for thousands of other genes. Transcriptional profile of CPCs exhibited greater degree of similarity throughout the cultured population relative to freshly isolated cells. Findings were validated by comparative analyses using scRNA-Seq datasets of various cell types generated by multiple scRNA-Seq technology. Increased transcriptome diversity and decreased population heterogeneity in the cultured cell population may help account for reported outcomes associated with experimental and clinical use of CPCs for treatment of myocardial injury.
用于心肌疾病的再生治疗方法通常涉及体外扩增的干细胞的递送。先前的研究表明，细胞培养条件会影响细胞的功能和表型特征，但从新鲜分离的群体中获得的培养细胞的关系和培养群体的异质性仍未得到很好的定义。体外扩增细胞的功能和表型特征将决定疾病干预治疗的结果，因此需要描述体外扩增对分离的干细胞群体的影响。单细胞 RNA 测序分析（scRNA-Seq）用于确定培养扩增对成人心肌祖细胞（CPC）的影响，以及与其他细胞群体的关系。生物信息学分析表明，在新鲜分离的细胞中表达的身份标记基因在培养的 CPC 中变得无法检测到，而数千个其他基因的表达水平则很低。与新鲜分离的细胞相比，CPC 的转录谱在整个培养群体中表现出更高的相似性。通过使用多种 scRNA-Seq 技术生成的各种细胞类型的 scRNA-Seq 数据集进行的比较分析验证了这些发现。培养细胞群体中转录组多样性的增加和群体异质性的降低可能有助于解释与 CPC 用于治疗心肌损伤的实验和临床应用相关的报告结果。
