Spatial distribution and initial changes of SSEA-1 and other cell adhesion-related molecules on mouse embryonic stem cells before and during differentiation
Spatial distribution and initial changes of SSEA-1 and other cell adhesion-related molecules on mouse embryonic stem cells before and during differentiation
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DOI：
10.1369/jhc.3a6241.2004
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发表时间：
2004-11-01
影响因子：
3.2
通讯作者：
Sasaki, K
中科院分区：
文献类型：
--
作者：
Cui, L;Johkura, K;Sasaki, K
我们研究了细胞粘附相关分子（CAM）在小鼠胚胎干细胞（ES）中的分布以及分化前和分化过程中细胞表面的空间分布。通过免疫组织化学和免疫扫描电镜，ES细胞集落中未分化细胞中SSEA-1、PECAM-1和ICAM-1的细胞-细胞异质性是明显的，支持流式细胞术的结果。相反，大多数未分化的ES细胞强烈表达CD 9。SSEA-1主要定位于低突起和微绒毛的边缘，形成3-20个颗粒的簇或线性阵列。PECAM-1和ICAM-1随机定位于游离细胞表面，而CD 9优先定位于微绒毛或突起，特别是在细胞周边。磁性细胞分选（MACS）的SSEA-1（+）级分和SSEA-1（-）级分在接种后均形成未分化的集落。流式细胞术显示，这些群体再次分别恢复到具有混合表型的培养物。视黄酸诱导的分化下调所有CAM的表达。免疫扫描电镜显示，SSEA-1在分化的ES细胞中减少，但仍存在一些聚集。我们的研究结果有助于阐明这些分子在ES细胞维持和分化中的意义，并表明细胞表面抗原可能有助于定义未分化和分化ES细胞的表型。
We examined the distribution of cell adhesion-related molecules (CAMs) among mouse embryonic stem (ES) cells and the spatial distribution on cell surfaces before and during differentiation. The cell-cell heterogeneity of SSEA-1, PECAM-1, and ICAM-1 among the undifferentiated cells in the ES cell colonies was evident by immunohistochemistry and immuno-SEM, supporting the flow cytometry findings. in contrast, most undifferentiated ES cells strongly expressed CD9. SSEA-1 was located preferentially on the edge of low protuberances and microvilli and formed clusters or linear arrays of 3-20 particles. PECAM-1 and ICAM-1 were randomly localized on the free cell surfaces, whereas CD9 was preferentially localized on the microvilli or protuberances, especially in the cell periphery. Both the SSEA-1(+) fraction and the SSEA-1(-) fraction of magnetic cell sorting (MACS) formed undifferentiated colonies after plating. Flow cytometry showed that these populations reverted separately again to a culture with a mixed phenotype. Differentiation induced by retinoic acid downregulated the expression of all CAMs. Immuno-SEM showed decreases of SSEA-1 in the differentiated ES cells, although some clustering still remained. Our findings help to elucidate the significance of these molecules in ES cell maintenance and differentiation and suggest that cell surface antigens may be useful for defining the phenotype of undifferentiated and differentiated ES cells.