Institute of Genetic Medicine, International Centre for Life, Newcastle University, Newcastle upon Tyne, NE1 3BZ, UK.
Stem Cell Rev Rep. 2013 Oct;9(5):731-9. doi: 10.1007/s12015-013-9440-9.
Cardiac side population (CSP) cells, defined by their ability to efflux the vital dye Hoechst 33342, have been identified as putative cardiac stem cells based on their potential to give rise to both cardiomyocytes and endothelial cells. The CSP phenotype relies on an active metabolic pathway and cell viability to identify a rare population of cells and therefore technical differences in the CSP staining protocol can lead to inconsistent results and discrepancies between studies. Here we describe an established protocol for CSP identification and have optimised a protocol for CSP analysis utilizing an automated cardiac digestion procedure using gentleMACs dissociation and Hoechst 33342 staining followed by dual wavelength flow cytometric analysis.
心脏侧群 (CSP) 细胞是指能够排出重要染料 Hoechst 33342 的细胞，它们被认为是潜在的心脏干细胞，因为它们具有产生心肌细胞和内皮细胞的潜力。CSP 表型依赖于活跃的代谢途径和细胞活力来识别罕见的细胞群体，因此 CSP 染色方案中的技术差异可能导致结果不一致，并且研究之间存在差异。在这里，我们描述了一种用于 CSP 鉴定的既定方案，并使用温和 MACs 分离和 Hoechst 33342 染色，然后进行双波长流式细胞术分析，优化了一种用于 CSP 分析的方案。
