Spontaneous hair follicle germ (HFG) formation in vitro, enabling the large-scale production of HFGs for regenerative medicine
Spontaneous hair follicle germ (HFG) formation in vitro, enabling the large-scale production of HFGs for regenerative medicine
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DOI：
10.1016/j.biomaterials.2017.10.056
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发表时间：
2018-02-01
期刊：
影响因子：
14
通讯作者：
Fukuda, Junji
中科院分区：
文献类型：
--
作者：
Kageyama, Tatsuto;Yoshimura, Chisa;Fukuda, Junji
毛囊形态发生是由毛囊胚（HFG）上皮层和间充质层之间的相互作用触发的。在这里，我们开发了一种通过细胞自组织在体外大规模制备HFGs的方法。我们将小鼠表皮细胞和小鼠/人间充质细胞悬浮混合，并将其接种在定制设计的阵列板的微孔中。在3天的培养期内，细胞最初形成随机分布的单细胞聚集体，然后在空间上彼此分离，表现出典型的HFG形态学特征。这些自我分选的毛囊胚芽（ssHFGs）被证明是能够有效的毛囊和轴后，皮内移植到裸鼠背部的生成。这一发现有助于在由透氧硅树脂制成的微孔阵列芯片中大规模制备约5000个ssHFG。我们证明了通过硅胶片底部的氧气供应的完整性对于实现ssHFG形成和随后的毛干生成至关重要。最后，将芯片上的空间对齐的ssHFGs封装到水凝胶中，同时移植到裸鼠的背部皮肤中以保留它们的介入空间，从而产生空间对齐的毛囊。这种简单的ssHFG制备方法是改善当前毛发再生医学技术的一种有前途的策略。(c)2017爱思唯尔有限公司版权所有
Hair follicle morphogenesis is triggered by reciprocal interactions between hair follicle germ (HFG) epithelial and mesenchymal layers. Here, we developed a method for large-scale preparation of HFGs in vitro via self-organization of cells. We mixed mouse epidermal and mouse/human mesenchymal cells in suspension and seeded them in microwells of a custom-designed array plate. Over a 3-day culture period, cells initially formed a randomly distributed single cell aggregate and then spatially separated from each other, exhibiting typical HFG morphological features. These self-sorted hair follicle germs (ssHFGs) were shown to be capable of efficient hair-follicle and shaft generation upon intracutaneous transplantation into the backs of nude mice. This finding facilitated the large-scale preparation of approximately 5000 ssHFGs in a microwell-array chip made of oxygen-permeable silicone. We demonstrated that the integrity of the oxygen supply through the bottom of the silicone chip was crucial to enabling both ssHFG formation and subsequent hair shaft generation. Finally, spatially aligned ssHFGs on the chip were encapsulated into a hydrogel and simultaneously transplanted into the back skin of nude mice to preserve their intervening spaces, resulting in spatially aligned hair follicle generation. This simple ssHFG preparation approach is a promising strategy for improving current hair-regenerative medicine techniques. (c) 2017 Elsevier Ltd. All rights reserved.