Current treatment options for uterine fibroids are limited to hormonal manipulation or surgical intervention. We aimed to develop an in vitro model to mirror collagen deposition and extracellular matrix (ECM) formation, the principal features of uterine fibroids, to enable testing of novel therapeutics. Macromolecular crowding with Ficoll 400 and Ficoll 70 in cultures of human uterine myometrial smooth muscle cells containing ascorbic acid, provided the basis for this model. These culture conditions mimic the ‘crowded’ nature of the in vivo extracellular environment by incorporating neutral, space-filling macromolecules into conventional cell cultures. This method of culture facilitates appropriate ECM deposition, thus closely representing the in vivo fibrotic phenotype of uterine fibroids. Macromolecular crowding in Ficoll cultures containing ascorbic acid reduced myometrial smooth muscle cell proliferation and promoted collagen production. Under these conditions, collagen was processed for extracellular deposition as demonstrated by C-propeptide cleavage from secreted procollagen. The fibrosis marker activin was increased relative to its natural inhibitor, follistatin, in crowded culture conditions while addition of exogenous follistatin reduced collagen (Col1A1) gene expression. This in vitro model represents a promising development for the testing of therapeutic interventions for uterine fibroids. However, it does not recapitulate the full in vivo pathology which can include specific genetic and epigenetic alterations that have not been identified in the myometrial smooth muscle (hTERT-HM) cell line. Following screening of potential therapeutics using the model, the most promising compounds will require further assessment in the context of individual subjects including those with genetic changes implicated in fibroid pathogenesis.

中文翻译：

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肌瘤病理学建模：子宫平滑肌细胞大分子拥挤模型的开发和操作以改变细胞外基质沉积。

目前子宫肌瘤的治疗选择仅限于激素控制或手术干预。我们旨在开发一种体外模型，以反映胶原蛋白沉积和细胞外基质 (ECM) 形成（子宫肌瘤的主要特征），从而能够测试新疗法。Ficoll 400 和 Ficoll 70 在含有抗坏血酸的人子宫肌层平滑肌细胞培养物中的大分子拥挤为该模型提供了基础。这些培养条件通过将中性的、充满空间的大分子纳入常规细胞培养物中来模拟体内细胞外环境的“拥挤”性质。这种培养方法有利于适当的 ECM 沉积，从而密切代表体内子宫肌瘤的纤维化表型。含有抗坏血酸的 Ficoll 培养物中的大分子拥挤减少了子宫肌层平滑肌细胞增殖并促进了胶原蛋白的产生。在这些条件下，胶原蛋白被加工用于细胞外沉积，如从分泌的原胶原蛋白切割 C-前肽所证明的。在拥挤的培养条件下，纤维化标志物激活素相对于其天然抑制剂卵泡抑素增加，而外源性卵泡抑素的添加减少了胶原蛋白 (Col1A1) 基因表达。这种体外模型代表了子宫肌瘤治疗干预测试的有希望的发展。然而，它并没有概括完整的体内病理学可能包括尚未在子宫肌层平滑肌 (hTERT-HM) 细胞系中鉴定的特定遗传和表观遗传改变。在使用该模型筛选潜在的治疗方法后，最有希望的化合物将需要在个体受试者的背景下进一步评估，包括那些与肌瘤发病机制有关的遗传变化。