Ma Sen, Wang Lamei, Zong Bo, Wang Ying, Wang Xiaolong, Shi Yinghua, Yang Yuxin, Chen Yulin
College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China.
Henan Key Laboratory of Innovation and Utilization of Grassland Resources, Zhengzhou 450002, China.
Animals (Basel). 2020 Aug 12;10(8):1400. doi: 10.3390/ani10081400.
A functional interpretation of filtered candidates and predicted regulatory pathways related to cashmere growth from sequencing trials needs available cell models, especially for hair matrix cells (HMCs), whose continual proliferation and differentiation result in rapid hair growth. To fulfill such goals, we herein obtained primary goat HMCs via a microdissection-based method; optimized the selection of the culture medium and coating substances for better cell maintenance; and exemplified their usefulness through examining the effects of calcium and all-trans retinoic acid (ATRA) on cells using immunoblotting, flow cytometry, and other techniques. As a result, we successfully acquired primary and passaged goat HMCs with typical keratinocyte morphology. Calcium-free RPMI (Roswell Park Memorial Institute) 1640 and MEM (minimum Eagle's medium) outperformed normal DMEM/F12 (Dulbecco's modified Eagle's medium/Nutrient Mixture F-12) on long-term cell maintenance, whereas serum-free media K-SFM and EpiLife failed to support cell growth. HMCs differed molecularly and morphologically from their neighbor dermal papilla cells on expressions of feature genes, such as , and on characteristic keratinocyte-like appearances versus fibroblast shapes, respectively. Higher calcium concentrations significantly stimulated the expression of the genes (e.g., and ) involved in keratinocyte differentiation and, promoted cell proliferation. Moreover, 10 M ATRA obviously boosted goat HMC expansions and changed their cell cycle distributions compared to the controls. Our study shines a light on researches exploring the mechanisms underlying the growth of cashmere.
对来自测序试验的与羊绒生长相关的过滤候选基因和预测调控途径进行功能解读需要可用的细胞模型,尤其是对于毛母细胞(HMCs),其持续增殖和分化导致毛发快速生长。为实现这些目标,我们在此通过基于显微切割的方法获得了原代山羊HMCs;优化了培养基和包被物质的选择以更好地维持细胞;并通过免疫印迹、流式细胞术和其他技术检测钙和全反式维甲酸(ATRA)对细胞的影响来例证它们的实用性。结果,我们成功获得了具有典型角质形成细胞形态的原代和传代山羊HMCs。无钙的RPMI(罗斯威尔公园纪念研究所)1640和MEM(伊格尔最低限度培养基)在长期细胞维持方面优于普通的DMEM/F12(杜尔贝科改良伊格尔培养基/营养混合物F-12),而无血清培养基K-SFM和EpiLife不能支持细胞生长。HMCs在特征基因表达上,以及在分别呈现特征性角质形成细胞样外观与成纤维细胞形状方面,在分子和形态上与其相邻的真皮乳头细胞不同。较高的钙浓度显著刺激参与角质形成细胞分化的基因(如 和 )的表达,并促进细胞增殖。此外,与对照相比,10 μM ATRA明显促进了山羊HMCs的扩增并改变了它们的细胞周期分布。我们的研究为探索羊绒生长潜在机制的研究提供了线索。