Epithelial cells derived from human embryonic stem cells display p16INK4A senescence, hypermotility, and differentiation properties shared by many P63+ somatic cell types.
Epithelial cells derived from human embryonic stem cells display p16INK4A senescence, hypermotility, and differentiation properties shared by many P63+ somatic cell types.
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DOI：
10.1002/stem.64
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发表时间：
2009-06
期刊：
影响因子：
5.2
通讯作者：
Rheinwald, James G.
中科院分区：
文献类型：
--
作者：
Dabelsteen, Sally;Hercule, Paula;Barron, Patricia;Rice, Meghan;Dorsainville, Gregory;Rheinwald, James G.
人胚胎干细胞（hES）可以产生表达p63、K14和外皮蛋白的细胞，这些细胞被认为是角质形成细胞。虽然这些hES衍生的角质形成细胞样（hESderK）细胞在对于正常组织衍生的角质形成细胞最佳的成纤维细胞饲养系统中培养时形成上皮样集落，但是它们具有非常短的复制寿命，除非被工程化以表达HPV 16 E6 E7。我们在这里报告，hESderK细胞经历衰老与p16 INK 4A的表达，无关的端粒状态。转导表达bmi 1，p16 INK 4A/p14 ARF基因座的阻遏物，赋予hESderK细胞和角质形成细胞显著延长的寿命。当暴露于转化生长因子β或层粘连蛋白-332的不完全加工形式时，我们研究的三种寿命延长或永生化的hESderK系变得定向运动过度，这是先前在角质形成细胞中表征的伤口愈合和侵袭反应。在器官型培养中，hESderK细胞分层并表达外皮蛋白和K10，体内表皮角质形成细胞也是如此。然而，它们的生长要求不如角质形成细胞严格。然后，我们将比较扩展到内胚层来源的p63+/K14+尿路上皮和气管支气管上皮细胞。这些细胞类型的原代细胞和永生化细胞系具有与角质形成细胞相似的生长要求和高运动性反应，并且通过工程化以表达端粒酶催化亚基（TERT），bmi 1表达促进了它们的永生化。在器官型培养，他们分层，并表现出鳞状化生，表达外皮蛋白和K10。因此，hESderK细胞被证明是不同于所有三个正常的p63+细胞类型测试。这些结果表明，hESderK细胞不能最终确定为角质形成细胞，甚至作为外胚层细胞，但可能代表一个不完整的形式，或偏离，正常的p63+谱系发展。
Human embryonic stem (hES) cells can generate cells expressing p63, K14, and involucrin, which have been proposed to be keratinocytes. Although these hES-derived, keratinocyte-like (hESderK) cells form epithelioid colonies when cultured in a fibroblast feeder system optimal for normal tissue-derived keratinocytes, they have a very short replicative lifespan unless engineered to express HPV16 E6E7. We report here that hESderK cells undergo senescence associated with p16INK4A expression, unrelated to telomere status. Transduction to express bmi1, a repressor of the p16INK4A/p14ARF locus, conferred upon hESderK cells and keratinocytes a substantially extended lifespan. When exposed to transforming growth factor beta or to an incompletely processed form of Laminin-332, three lifespan-extended or immortalized hESderK lines that we studied became directionally hypermotile, a wound healing and invasion response previously characterized in keratinocytes. In organotypic culture, hESderK cells stratified and expressed involucrin and K10, as do epidermal keratinocytes in vivo. However, their growth requirements were less stringent than keratinocytes. We then extended the comparison to endoderm-derived, p63+/K14+ urothelial and tracheobronchial epithelial cells. Primary and immortalized lines of these cell types had growth requirements and hypermotility responses similar to keratinocytes and bmi1 expression facilitated their immortalization by engineering to express the catalytic subunit of telomerase (TERT). In organotypic culture, they stratified and exhibited squamous metaplasia, expressing involucrin and K10. Thus, hESderK cells proved to be distinct from all three normal p63+ cell types tested. These results indicate that hESderK cells cannot be identified conclusively as keratinocytes or even as ectodermal cells, but may represent an incomplete form of, or deviation from, normal p63+ lineage development.
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影响因子：
6.5
作者：
HORMIA, M;FALKMARZILLIER, J;QUARANTA, V
通讯作者：
QUARANTA, V
影响因子：
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作者：
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通讯作者：
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影响因子：
4.3
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Rouleau, Matthieu
影响因子：
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作者：
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通讯作者：
Benvenisty, N
影响因子：
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作者：
Koster, MI;Kim, S;Roop, DR
通讯作者：
Roop, DR