Dynamic Culture Substrates That Mimic the Topography of the Epidermal-Dermal Junction.
Dynamic Culture Substrates That Mimic the Topography of the Epidermal-Dermal Junction.
复制标题
DOI：
10.1089/ten.tea.2018.0125
复制
发表时间：
2019-03
影响因子：
--
通讯作者：
Watt FM
中科院分区：
文献类型：
--
作者：
Helling AL;Viswanathan P;Cheliotis KS;Mobasseri SA;Yang Y;El Haj AJ;Watt FM
人皮肤的表皮层和真皮层之间的连接处呈波浪状，波浪的宽度和深度随年龄和疾病而变化。我们先前表明，当原代人表皮角质形成细胞接种在涂有胶原蛋白的波浪状静态聚二甲基硅氧烷（PDMS）弹性体基底上时，干细胞、分化细胞和增殖细胞会根据来自底层基底的信号而形成特定模式。为了研究这种模式是如何随时间形成的，我们现在创建了一个动态模型，在该模型中，将涂有胶原蛋白的聚（D,L - 丙交酯 - 乙交酯）（PLGA）膜放置在含有不同直径和间距的圆形孔的聚酰亚胺片上。当施加真空时，膜会被诱导产生波浪状起伏，起伏的高度取决于所施加的压力和孔的大小。我们观察到在施加真空后的48小时内，具有高水平β1整合素表达（一种干细胞标记）的细胞在起伏的底部聚集。分化的外皮蛋白阳性细胞没有聚集；然而，具有高E - 钙黏蛋白表达和核YAP的细胞出现了聚集。Rho激酶抑制导致聚集消失，这表明Rho家族成员在该过程中起作用。
在人皮肤中，表皮和真皮之间的连接处呈波浪状。表皮干细胞根据其相对于这些波浪状结构的位置形成特定模式。在这里我们描述了一种装置，其中表皮细胞在涂有胶原蛋白的聚（D,L - 丙交酯 - 乙交酯）（PLGA）膜上培养。当施加真空时，膜被诱导产生波浪状起伏。干细胞会因真空而聚集，而分化细胞则不会。Rho激酶抑制导致聚集消失，这表明Rho家族成员在该过程中起作用。这个动态平台是研究皮肤随年龄和疾病变化的一种新工具。
The junction between the epidermal and dermal layers of human skin undulates, the width and depth of the undulations varying with age and disease. We previously showed that when primary human epidermal keratinocytes are seeded on collagen-coated undulating static polydimethylsiloxane (PDMS) elastomer substrates, the stem cells, differentiated cells, and proliferating cells become patterned in response to cues from the underlying substrate. To investigate how patterning occurs over time, we have now created a dynamic model, in which a collagen-coated poly(d,l-lactide-co-glycolide) (PLGA) membrane is placed over a polyimide sheet containing circular holes, differing in diameter and spacing. When a vacuum is applied the membrane is induced to undulate, the heights of the undulations depending on the pressure applied and the size of the holes. We observed clustering of cells with high levels of β1 integrin expression, a stem cell marker, in the base of the undulations within 48 h of applying the vacuum. Differentiating involucrin-positive cells did not cluster; however, there was clustering of cells with high E-cadherin expression and nuclear YAP. Rho kinase inhibition resulted in loss of clustering, suggesting a role for Rho family members in the process. In human skin the junction between the epidermis and dermis undulates. Epidermal stem cells pattern according to their position relative to those undulations. Here we describe a rig in which epidermal cells are cultured on a collagen-coated poly(d,l-lactide-co-glycolide) (PLGA) membrane. When a vacuum is applied the membrane is induced to undulate. Stem cells cluster in response to the vacuum, whereas differentiating cells do not. Rho kinase inhibition results in loss of clustering, suggesting a role for Rho family members in the process. This dynamic platform is a new tool for investigating changes in the skin with age and disease.