Identification and characterization of side population cells in embryonic stem cell cultures.
Identification and characterization of side population cells in embryonic stem cell cultures.
复制标题
胚胎干细胞培养物中侧群细胞的鉴定和表征。
DOI：
10.1089/scd.2008.0391
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发表时间：
2009
影响因子：
4
通讯作者：
Goodell,MargaretA
中科院分区：
文献类型：
--
作者：
Vieyra,DiegoS;Rosen,Allison;Goodell,MargaretA
胚胎干细胞（ESC）的标记和功能异质性已被描述。这一特性与类似于胚胎多能细胞谱系的 ESC 亚群的存在相关。侧群 (SP) 细胞所表现出的 Hoechst (Ho) 外排能力已被证明可作为识别多种组织中的多能干细胞的标记物。在这里，我们报告来自不同 ESC 系的培养物一致地显示出 SP 群体，该群体表现出未分化 ESC 的抗原、独特的药物流出特性以及 ABC 转运蛋白、内细胞团 (ICM) 和外胚层基因的表达模式，这将其与非 SP ESC 部分区分开来。该 SP 群体包含在胚状体和畸胎瘤检测中分化为外胚层、中胚层和内胚层的多能细胞。此外，纯化的 SP 细胞有效整合到发育中的桑葚中并有助于 ICM。在标准ESC培养条件下，SP和非SP群体表现出相互转化的能力；然而，这些分数之间建立了平衡。使用为 SP ESC 定制的方案，我们报告可以在植入周围囊胚的 ICM 中识别出具有相似外排特性的细胞。我们的结果表明，ESC 显示出 SP 标记的异质性，并且这些培养物的 SP 群体包含在表型和功能上类似于植入周围胚胎的外排活性 ICM 细胞的细胞。我们的观察结果表明 SP 表型参与 ESC 维持和早期胚胎发育，并支持 ESC 由处于动态平衡的不同表型和功能多能亚群组成的观点。
Marker and functional heterogeneity has been described for embryonic stem cells (ESCs). This property has been correlated with the presence of ESC subpopulations resembling pluripotent cell lineages of the embryo. The ability to efflux Hoechst (Ho) displayed by side population (SP) cells has proven valuable as a marker to identify multipotent stem cells from a variety of tissues. Here we report that cultures from different ESC lines consistently show an SP population that displays antigens of undifferentiated ESCs, distinct drug efflux properties, and an expression pattern of ABC transporters, inner cell mass (ICM), and epiblast genes, which distinguish it from the non-SP ESC fraction. This SP population contains pluripotent cells that differentiate into ectoderm, mesoderm, and endoderm in embryoid body and teratoma assays. Further, purified SP cells efficiently integrate into developing morulae and contribute to ICM. Under standard ESC culture conditions, SP and non-SP populations display ability to convert into each other; however, an equilibrium establishes between these fractions. Using protocols customized for SP ESCs, we report that cells with similar efflux properties can be identified in the ICM of peri-implanted blastocysts. Our results indicate that ESCs display heterogeneity for the SP marker, and the SP population of these cultures contains cells that phenotypically and functionally resemble efflux-active ICM cells of the peri-implanted embryo. Our observations suggest an involvement of the SP phenotype in ESC maintenance and early embryo development, and support the idea that ESCs are composed of distinct phenotypic and functional pluripotent subpopulations in dynamic equilibrium.