Surface Tension Guided Hanging-Drop: Producing Controllable 3D Spheroid of High-Passaged Human Dermal Papilla Cells and Forming Inductive Microtissues for Hair-Follicle Regeneration
Surface Tension Guided Hanging-Drop: Producing Controllable 3D Spheroid of High-Passaged Human Dermal Papilla Cells and Forming Inductive Microtissues for Hair-Follicle Regeneration
复制标题
表面张力引导的悬滴：产生高传代人真皮乳头细胞的可控 3D 球体并形成用于毛囊再生的诱导微组织。
DOI：
10.1021/acsami.6b00202
复制
发表时间：
2016-03-09
影响因子：
9.5
通讯作者：
Xing, Malcolm
中科院分区：
文献类型：
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作者：
Lin, Bojie;Miao, Yong;Xing, Malcolm
人毛乳头（DP）细胞已被广泛研究时，在传统的单层生长。然而，由于与体内真实的三维（3D）环境的极大偏差，这些二维（2D）生长的细胞在传代过程中往往失去毛发诱导能力。因此，这些2D引起的问题已经促使开发新的3D培养技术，以产生具有合适模拟物的细胞微组织。悬滴法是基于表面张力技术和表面张力与重力场之间的相互作用，使液滴的收敛。本研究使用该技术在会聚滴中形成毛乳头细胞的细胞球体。它导致一个可控的三维球体模型的可扩展的制造诱导DP微组织。首先确定了培养高传代（P8）DP球状体的最佳条件。然后进行形态学、组织学和功能学研究。此外，毛发诱导标志物，包括碱性磷酸酶，α-平滑肌肌动蛋白和神经细胞粘附分子的表达也通过定量RT-PCR，免疫染色和免疫印迹分析。最后，将P8-DP微组织与新生小鼠表皮细胞（EPCs）共植入裸鼠体内。我们的研究结果表明，3D微组织的形成不仅赋予P8-DP微组织与初级DP的许多相似之处，而且赋予这些微组织增强的诱导毛囊（HF）新生的能力。该模型提供了一个潜在的阐明人类DP的天然生物学，也显示了可控的和可扩展的生产应用于未来的卵泡再生的诱导DP细胞的前景。
Human dermal papilla (DP) cells have been studied extensively when grown in the conventional monolayer. However, because of great deviation from the real in vivo three-dimensional (3D) environment, these two-dimensional (2D) grown cells tend to lose the hair-inducible capability during passaging. Hence, these 2D caused concerns have motivated the development of novel 3D culture techniques to produce cellular microtissues with suitable mimics. The hanging-drop approach is based on surface tension-based technique and the interaction between surface tension and gravity field that makes a convergence of liquid drops. This study used this technique in a converged drop to form cellular spheroids of dermal papilla cells. It leads to a controllable 3Dspheroid model for scalable fabrication of inductive DP microtissues. The optimal conditions for culturing high passaged (P8) DP spheroids were determined first. Then, the morphological, histological and functional studies were performed. In addition, expressions of hair-inductive markers including alkaline phosphatase, alpha-smooth muscle actin and neural cell adhesion molecule were also analyzed by quantitative RT-PCR, immunostaining and immunoblotting. Finally, P8-DP microtissues were coimplanted with newborn mouse epidermal cells (EPCs) into nude mice. Our results indicated that the formation of 3D microtissues not only endowed P8-DP microtissues many similarities to primary DP, but also confer these microtissues an enhanced ability to induce hair-follicle (HF) neogenesis in vivo. This model provides a potential to elucidate the native biology of human DP, and also shows the promising for the controllable and scalable production of inductive DP cells applied in future follicle regeneration.