Role of melanoma chondroitin sulphate proteoglycan in patterning stem cells in human interfollicular epidermis
Role of melanoma chondroitin sulphate proteoglycan in patterning stem cells in human interfollicular epidermis
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DOI：
10.1242/dev.00837
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发表时间：
2003-12-01
期刊：
影响因子：
4.6
通讯作者：
Watt, FM
中科院分区：
文献类型：
--
作者：
Legg, J;Jensen, UB;Watt, FM
人类毛囊间表皮被干细胞更新，这些干细胞以图案化的非随机分布聚集在基底层中。干细胞可以通过高表达β 1整合素和缺乏终末分化标记物的表达而与其他角质形成细胞区分开;它们在体内很少分裂，但在培养物中形成活跃生长的集落。在寻找其他干细胞标记物时，我们观察到黑色素瘤硫酸软骨素蛋白聚糖（MCSP）的异质性表皮表达。MCSP由具有最高β 1整联蛋白水平的角质形成细胞表达。在滤泡间表皮，表达仅限于非周期性细胞，并在文化中，自我更新的克隆。然而，基于MCSP和β 1整联蛋白表达的荧光激活细胞分选并不比单独分选β 1整联蛋白更富集克隆源性角质形成细胞。为了干扰内源性MCSP，我们逆转录病毒感染角质形成细胞与嵌合体的CD 8细胞外结构域和MCSP胞质结构域。CD 8/MCSP不影响角质形成细胞的增殖或分化，但角质形成细胞在分离的克隆或重建的表皮片的凝聚力大大降低。CD 8/MCSP导致干细胞后代分散而不分化。CD 8/MCSP没有改变角质形成细胞的运动，但干扰钙粘蛋白介导的细胞-细胞粘附和皮质肌动蛋白细胞骨架，可以通过抑制Rho模拟的效果。我们的结论是，MCSP是一种新的标记表皮干细胞，有助于他们的模式分布，促进干细胞聚集。
Human interfollicular epidermis is renewed by stem cells that are clustered in the basal layer in a patterned, non-random distribution. Stem cells can be distinguished from other keratinocytes by high expression of beta1 integrins and lack of expression of terminal differentiation markers; they divide infrequently in vivo but form actively growing colonies in culture. In a search for additional stem cell markers, we observed heterogeneous epidermal expression of melanoma chondroitin sulphate proteoglycan (MCSP). MCSP was expressed by those keratinocytes with the highest beta1 integrin levels. In interfollicular epidermis, expression was confined to non-cycling cells and, in culture, to self-renewing clones. However, fluorescence-activated cell sorting on the basis of MCSP and beta1 integrin expression gave no more enrichment for clonogenic keratinocytes than sorting for beta1 integrins alone. To interfere with endogenous MCSP, we retrovirally infected keratinocytes with a chimera of the CD8 extracellular domain and the MCSP cytoplasmic domain. CD8/MCSP did not affect keratinocyte proliferation or differentiation but the cohesiveness of keratinocytes in isolated clones or reconstituted epidermal sheets was greatly reduced. CD8/MCSP caused stem cell progeny to scatter without differentiating. CD8/MCSP did not alter keratinocyte motility but disturbed cadherin-mediated cell-cell adhesion and the cortical actin cytoskeleton, effects that could be mimicked by inhibiting Rho. We conclude that MCSP is a novel marker for epidermal stem cells that contributes to their patterned distribution by promoting stem cell clustering.