Dermal papilla cells induce keratinocyte tubulogenesis in culture
Dermal papilla cells induce keratinocyte tubulogenesis in culture
复制标题
DOI：
10.1007/s00418-010-0691-0
复制
发表时间：
2010-05-01
影响因子：
2.3
通讯作者：
Terskikh, Vasily V.
中科院分区：
文献类型：
--
作者：
Chermnykh, Elina S.;Vorotelyak, Ekaterina A.;Terskikh, Vasily V.
许多研究报道了真皮乳头（DP）细胞诱导毛发生长的能力。然而，人们对毛囊发育的早期阶段和控制这一过程的信号知之甚少。因此，体外模型可能是研究上皮-间充质相互作用和表皮形态发生早期阶段的方便系统，特别是在人类中。为了研究DP细胞在人头皮毛囊表皮形态发生中的作用，我们改进了从人头皮毛囊中分离DP细胞的方法，并建立了表皮形态发生的三维模型。分离的DP细胞能够向成脂和成骨方向分化，并在培养过程中保持碱性磷酸酶（AP）活性达7代。DP细胞在体外三维模型中能诱导出小管样结构，并能重组胶原基质。由于多次传代后毛囊诱导能力和生长活性下降，DP细胞的长时间培养一直是一个大问题。为了解决这一问题，我们通过转染人端粒酶逆转录酶cDNA （hTERT）实现了DP细胞的永生化。永生化DP-hTERT细胞保留AP活性，表现出较低的成骨分化能力。从活跃增殖细胞中收集的条件培养基以及DP-hTERT细胞本身在角化细胞长时间培养后能够诱导小管形成。
The ability of dermal papilla (DP) cells to induce hair growth was reported in many studies. However, early stages of hair follicle development and signals that govern this process are poorly understood. Therefore, an in vitro model may be a convenient system to study epithelial-mesenchymal interactions and early stages of epidermal morphogenesis, especially in humans. To investigate the role of DP cells in epidermal morphogenesis we modified the method of isolation of DP cells from hair follicle of human scalp and developed the three-dimensional model of epidermal morphogenesis. Isolated DP cells were able to differentiate in adipogenic and osteogenic directions and retained activity of alkaline phosphatase (AP) for seven passages in culture. DP cells were able to induce tubule-like structures in three-dimensional model in vitro and to reorganize collagen matrix. Prolonged cultivation of DP cells has been a big problem because of the loss of hair follicle-inducing ability and growth activity after several passages. To solve this problem we immortalized DP cells by the transfection of the human telomerase reverse transcriptase cDNA (hTERT). Immortalized DP-hTERT cells retained AP activity and demonstrated low ability to osteogenic differentiation. The conditioned medium collected from actively proliferated cells as well as DP-hTERT cells themselves were capable to induce tubulogenesis after prolonged keratinocyte cultivation.