PDF(Pubmed)
Abstract：
This study investigated the three-dimensional (3D) cellular interactions and tunneling nanotubes (TNTs) during fetal mouse skin regeneration on embryonic days 13 (E13) and 15 (E15). We aimed to understand spatial relationships among cell types involved in skin regeneration and assess the potential role of TNTs. Full-thickness skin incisions were performed in E13 and E15 embryos. Wound sites were collected, embedded in epoxy resin, processed for 3D reconstruction (1 μm thickness sections), and subjected to whole-mount immunostaining. We conducted in vitro co-culture experiments with fetal macrophages and fibroblasts to observe TNT formation. To assess the effect of TNTs on skin regeneration, an inhibiting agent (cytochalasin B) was administered to amniotic fluid. Results revealed that E13 epidermal keratinocytes interacted with dermal fibroblasts and macrophages, facilitating skin regrowth. TNT structures were observed at the E13-cell wound sites, among macrophages, and between macrophages and fibroblasts, confirmed through in vitro co-culture experiments. In vitro and utero cytochalasin B administration hindered those formation and inefficient skin texture regeneration at E13 wound sites. This emphasizes the necessity of 3D cellular interactions between epidermal and dermal cells during skin regeneration in mouse embryos at E13. The prevalence of TNT structures indicated their involvement in achieving complete skin texture restoration.
摘要：
这项研究调查了胚胎第13天（E13）和第15天（E15）的胎儿小鼠皮肤再生过程中的三维（3D）细胞相互作用和隧道纳米管（TNTs）。我们旨在了解参与皮肤再生的细胞类型之间的空间关系，并评估TNTs的潜在作用。在E13和E15胚胎中进行全厚度皮肤切口。收集伤口部位，嵌入环氧树脂中，处理用于三维重建(厚度为1μm的切片)，并进行整体免疫染色。我们与胎儿巨噬细胞和成纤维细胞进行了体外共培养实验，以观察TNT的形成。为了评估TNTs对皮肤再生的影响，将抑制剂（细胞松弛素B）给予羊水。结果显示，E13表皮角质形成细胞与真皮成纤维细胞和巨噬细胞相互作用，促进皮肤再生。在E13细胞伤口部位观察到TNT结构，在巨噬细胞中，在巨噬细胞和成纤维细胞之间，通过体外共培养实验证实。体外和子宫细胞松弛素B的施用阻碍了E13伤口部位的这些形成和低效的皮肤质地再生。这强调了E13小鼠胚胎皮肤再生过程中表皮和真皮细胞之间3D细胞相互作用的必要性。TNT结构的流行表明它们参与实现完整的皮肤纹理恢复。
