Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China.
Department of Burns, Plastic, Cosmetology and Wound Repair Surgery, Guangzhou First People's Hospital, Guangzhou, Guangdong 510180, China.
Acta Biomater. 2023 Jul 15;165:31-49. doi: 10.1016/j.actbio.2022.11.004. Epub 2022 Nov 5.
The restoration of hair-inductive potential in human dermal papilla cells (hDPCs) is a tremendous challenge for hair regeneration. Much of the research thus far has indicated that three-dimensional (3-D) culture shows improved efficacy in hair follicle (HF) neogenesis. However, mature HF cannot regenerate in an incomplete microenvironment. This study developed an optimized 3-D co-culture system to restore the hair-inductive characteristics of hDPCs by mimicking the in-vivo microenvironment. As a result, Matrigel-encapsulated hDPCs spontaneously formed into hDPC aggregates (hDPAs), which exhibited better activity, higher proliferation rates, and less apoptosis and hypoxia than the ultra-low attachment culture. Interestingly, the co-culture with the hair matrix cells and dermal sheath cup cells further enhanced the expression of hair regeneration-related genes of hDPAs compared to conditioned medium and improved mature HF induction. In addition, these hDPAs with higher hair inductivity could be produced on a large scale and easily separated for gene expression detection. Finally, the mRNA sequencing, PCR, and WB results showed that the co-culture biomimetic microenvironment stimulated the canonical Wnt signaling pathway and inhibited the BMP signaling pathway. Thus, this co-culture system will provide a reliable platform that allows high-throughput culture, testing, and harvesting of hDPAs for HF tissue engineering. STATEMENT OF SIGNIFICANCE: Extensive hair loss continues to be difficult to treat and causes significant patient morbidity. Hair follicle (HF) tissue engineering may seem to be a way out. However, the absence of the in-vivo microenvironment fails to regenerate mature hairs. This study systematically described a biomimetic co-culture approach to generate better quality human dermal papilla cell aggregates (hDPAs) with improved hair inductive properties, which can be further used for HF tissue engineering. The hDPC microenvironment was reprogrammed through the controllable formation of self-assembled organoids in Matrigel and the tri-culture with hair matrix cells and dermal sheath cup cells. This work indicates that the production of hDPAs could be readily scaled, in theory for large-scale assays, analyses, or therapeutic applications.
毛发诱导潜能在人类真皮乳头细胞（hDPCs）中的恢复是毛发再生的巨大挑战。迄今为止，大量研究表明，三维（3-D）培养在毛囊（HF）新生方面显示出更好的效果。然而，成熟的 HF 不能在不完全的微环境中再生。本研究通过模拟体内微环境，开发了一种优化的 3-D 共培养系统，以恢复 hDPC 的毛发诱导特性。结果，Matrigel 包被的 hDPCs 自发形成 hDPC 聚集体（hDPAs），与超低附着培养相比，其活性更高、增殖率更高、凋亡和缺氧更少。有趣的是，与条件培养基相比，与毛基质细胞和真皮鞘杯细胞的共培养进一步增强了 hDPAs 中与毛发再生相关基因的表达，并提高了成熟 HF 的诱导。此外，这些具有更高毛发诱导性的 hDPAs 可以大规模生产，并易于分离进行基因表达检测。最后，mRNA 测序、PCR 和 WB 结果表明，共培养仿生微环境刺激了经典 Wnt 信号通路，抑制了 BMP 信号通路。因此，这种共培养系统将为 HF 组织工程提供一个可靠的平台，允许高通量培养、测试和收获 hDPAs。
广泛的脱发仍然难以治疗，并导致患者严重发病。毛囊（HF）组织工程似乎是一种出路。然而，缺乏体内微环境无法再生成熟的毛发。本研究系统地描述了一种仿生共培养方法，以产生具有改善毛发诱导特性的更好质量的人真皮乳头细胞聚集体（hDPAs），可进一步用于 HF 组织工程。通过在 Matrigel 中可控形成自组装类器官和与毛基质细胞和真皮鞘杯细胞的三培养，重新编程 hDPC 微环境。这项工作表明，hDPAs 的生产可以在理论上进行大规模放大，用于大规模测定、分析或治疗应用。
