Martier Ashley T, Maurice Yasmin V, Conrad K Michael, Mauvais-Jarvis Franck, Mondrinos Mark J
Department of Biomedical Engineering, Tulane University School of Science & Engineering, New Orleans, LA, USA.
Tulane Center for Excellence in Sex-based Biology and Medicine, New Orleans, LA, USA.
bioRxiv. 2023 Jul 25:2023.07.23.550236. doi: 10.1101/2023.07.23.550236.
Progress toward the development of sex-specific tissue engineered systems has been hampered by the lack of research efforts to define the effects of sex-specific hormone concentrations on relevant human cell types. Here, we investigated the effects of defined concentrations of estradiol (E2) and dihydrotestosterone (DHT) on primary human dermal and lung fibroblasts (HDF and HLF), and human umbilical vein endothelial cells (HUVEC) from female (XX) and male (XY) donors in both 2D expansion cultures and 3D stromal vascular tissues. Sex-matched E2 and DHT stimulation in 2D expansion cultures significantly increased the proliferation index, mitochondrial membrane potential, and the expression of genes associated with bioenergetics (Na+/K+ ATPase, somatic cytochrome C) and beneficial stress responses (chaperonin) in all cell types tested. Notably, cross sex hormone stimulation, i.e., DHT treatment of XX cells in the absence of E2 and E2 stimulation of XY cells in the absence of DHT, decreased bioenergetic capacity and inhibited cell proliferation. We used a microengineered 3D vasculogenesis assay to assess hormone effects on tissue scale morphogenesis. E2 increased metrics of vascular network complexity compared to vehicle in XX tissues. Conversely, and in line with results from 2D expansion cultures, E2 potently inhibited vasculogenesis compared to vehicle in XY tissues. DHT did not significantly alter vasculogenesis in XX or XY tissues but increased the number of non-participating endothelial cells in both sexes. This study establishes a scientific rationale and adaptable methods for using sex hormone stimulation to develop sex-specific culture systems.
由于缺乏研究来确定性别特异性激素浓度对相关人类细胞类型的影响，性别特异性组织工程系统的开发进展受到了阻碍。在此，我们研究了特定浓度的雌二醇（E2）和双氢睾酮（DHT）对来自女性（XX）和男性（XY）供体的原代人皮肤和肺成纤维细胞（HDF和HLF）以及人脐静脉内皮细胞（HUVEC）在二维扩增培养和三维基质血管组织中的影响。在二维扩增培养中，性别匹配的E2和DHT刺激显著提高了所有测试细胞类型的增殖指数、线粒体膜电位以及与生物能量学（Na+/K+ ATP酶、体细胞细胞色素C）和有益应激反应（伴侣蛋白）相关基因的表达。值得注意的是，跨性别激素刺激，即在没有E2的情况下用DHT处理XX细胞以及在没有DHT的情况下用E2刺激XY细胞，会降低生物能量能力并抑制细胞增殖。我们使用微工程三维血管生成试验来评估激素对组织规模形态发生的影响。与XX组织中的载体相比，E2增加了血管网络复杂性的指标。相反，与XY组织中的载体相比，E2强烈抑制血管生成，这与二维扩增培养的结果一致。DHT在XX或XY组织中均未显著改变血管生成，但增加了两性中未参与的内皮细胞数量。本研究为利用性激素刺激开发性别特异性培养系统建立了科学依据和适用方法。
