Marker succession during the development of keratinocytes from cultured human embryonic stem cells
Marker succession during the development of keratinocytes from cultured human embryonic stem cells
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DOI：
10.1073/pnas.0307226100
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发表时间：
2003-12-23
影响因子：
11.1
通讯作者：
Iuchi, S
中科院分区：
文献类型：
--
作者：
Green, H;Easley, K;Iuchi, S
注射到 scid 小鼠体内的人类胚胎干细胞产生含有分化体细胞组织的结节。从这种结节的胰蛋白酶消化细胞中，我们回收了可以在细胞培养物中生长的角质形成细胞。恢复方法足够灵敏，可以检测结节中形成的少量角质形成细胞，但出于分析目的，最好研究培养物中整个角质形成细胞谱系的发育。我们分析的原理是标记物的连续出现，包括对角质形成细胞具有相当特异性的转录因子（p63和basonuclin）和其最终状态特征的分化标记物（角蛋白14和外皮蛋白）。我们已经确定了细胞培养中单个胚状体角质形成细胞的时间和迁移依赖性发育过程中标记连续的顺序。在我们检查的标记中，p63 是最早出现在角质形成细胞谱系中的。后来标记的连续积累提供了最终角质形成细胞出现的增加的确定性。
Human embryonic stem cells injected into scid mice produce nodules containing differentiated somatic tissues. From the trypsinized cells of such a nodule, we have recovered keratinocytes that can be grown in cell culture. The method of recovery is sensitive enough to detect small numbers of keratinocytes formed in the nodule, but for purposes of analysis, it is preferable to study the development of the entire keratinocyte lineage in culture. The principle of our analysis is the successive appearance of markers, including transcription factors with considerable specificity for the keratinocyte (p63 and basonuclin) and differentiation markers characteristic of its final state (keratin 14 and involucrin). We have determined the order of marker succession during the time- and migration-dependent development of keratinocytes from single embryoid bodies in cell culture. Of the markers we have examined, p63 was the earliest to appear in the keratinocyte lineage. The successive accumulation of later markers provides increasing certainty of emergence of the definitive keratinocyte.