The Key Laboratory of Pathobiology, Ministry of Education, College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130000, P.R. China.
Department of Neurosurgery, China‑Japan Union Hospital of Jilin University, Changchun, Jilin 130000, P.R. China.
Mol Med Rep. 2021 May;23(5). doi: 10.3892/mmr.2021.11965. Epub 2021 Mar 24.
Hair loss, including alopecia, is a common and distressing problem for men and women, and as a result, there is considerable interest in developing treatments that can prevent or reverse hair loss. Dermal papillae closely interact with epidermal cells and play a key role during hair follicle induction and hair morphogenesis. As dermal papilla cells (DPCs) lose their hair‑inducing ability in monolayer cultures , it is difficult to obtain hair follicle structures following DPC transplantation . The present study aimed to explore culture conditions to maintain DPC characteristics using conditioned media (CM) from the supernatant of cultured HaCaT keratinocyte cells supplemented with other components. Initially, it was observed that during passaging of monolayer DPC cultures, the Wnt/β‑catenin pathway was repressed, while the TGF‑β/Smad pathway was activated, and that HaCaT cells cultivated in 1% fetal bovine serum had higher levels of expression of Wnt3a and Wnt10b compared with normal keratinocytes. Culturing of high‑passage (P7) DPCs in CM from HaCaT cells (HaCaT‑CM) actively stimulated cell proliferation and maintained Sox2 and Versican expression levels. Supplementation of HaCaT‑CM with SB431542 (SB, a TGF‑β receptor inhibitor), CHIR99021, (CHIR, a GSK3α/β inhibitor and activator of Wnt signaling) and platelet‑derived growth factor (PDGF)‑AA further increased the expression levels of Sox2, Versican and alkaline phosphatase (ALP) in P7 DPCs. Three‑dimensional culture of P7 DPCs using hanging drop cultures in HaCaT‑CM supplemented with SB, CHIR and PDGF‑AA resulted in larger cell aggregates and a further significant upregulation of Sox2, ALP and Versican expression levels. Taken together, these findings demonstrated that HaCaT‑CM supplemented with SB, CHIR and PDGF‑AA may preserve the hair‑inducing ability of high‑passage DPCs and may therefore be useful in reconstructing new hair follicles .
脱发，包括脱发症，是男性和女性常见且令人苦恼的问题，因此，人们对开发能够预防或逆转脱发的治疗方法产生了浓厚的兴趣。真皮乳头与表皮细胞密切相互作用，在毛囊诱导和毛发生成过程中发挥关键作用。由于真皮乳头细胞（DPC）在单层培养中丧失了诱导毛发的能力，因此在 DPC 移植后很难获得毛囊结构。本研究旨在通过添加其他成分的培养 HaCaT 角质形成细胞上清液的条件培养基（CM）来探索维持 DPC 特性的培养条件。最初观察到，在单层 DPC 培养物的传代过程中，Wnt/β-连环蛋白途径受到抑制，而 TGF-β/Smad 途径被激活，并且在 1%胎牛血清中培养的 HaCaT 细胞中，Wnt3a 和 Wnt10b 的表达水平高于正常角质形成细胞。在 HaCaT 细胞（HaCaT-CM）的 CM 中培养高传代（P7）DPC 可积极刺激细胞增殖，并维持 Sox2 和 Versican 的表达水平。用 SB431542（SB，TGF-β受体抑制剂）、CHIR99021（CHIR，GSK3α/β抑制剂和 Wnt 信号通路激活剂）和血小板衍生生长因子（PDGF）-AA 补充 HaCaT-CM 进一步增加了 P7 DPC 中 Sox2、Versican 和碱性磷酸酶（ALP）的表达水平。在补充有 SB、CHIR 和 PDGF-AA 的 HaCaT-CM 中通过悬滴培养对 P7 DPC 进行三维培养导致细胞聚集体更大，并进一步显著上调 Sox2、ALP 和 Versican 的表达水平。总之，这些发现表明，补充有 SB、CHIR 和 PDGF-AA 的 HaCaT-CM 可能保持高传代 DPC 的诱导毛发能力，因此可能有助于重建新的毛囊。
