Enhanced Differentiation Potential of Primary Human Endometrial Cells Cultured on 3D Scaffolds

Novel approaches for culturing primary human cells in vitro are increasingly needed to study cell and tissue physiology and to grow replacement tissue for regenerative medicine. Conventional 2D monolayer cultures of endometrial epithelial and stromal cells fail to replicate the complex 3D architecture of tissue. A fully synthetic scaffold that mimics the microenvironment of the human endometrium can ultimately provide a robust platform for investigating tissue physiology and, hence, take significant steps toward tackling female infertility and IVF failure. In this work, emulsion-templated porous polymers (known as polyHIPEs) were investigated as scaffolds for the culture of primary human endometrial epithelial and stromal cells (HEECs and HESCs). Infiltration of HEECs and HESCs into cell-seeded polyHIPE scaffolds was assessed by histological studies, and phenotype was confirmed by immunostaining. Confocal microscopy revealed that the morphology of HEECs and HESCs is representative of that found in vivo. RNA sequencing was used to investigate transcriptome differences between cells grown on polyHIPE scaffolds and in monolayer cultures. The differentiation status of HEECs and HESCs grown in polyHIPE scaffolds and in monolayer cultures was further evaluated by monitoring the expression of endometrial marker genes. Our observations suggest that a 3D cell culture model that could approximate native human endometrial architecture and function can be developed using tailored polyHIPE scaffolds.

体外培养原代人类细胞的新型方法，在细胞与组织生理学研究以及再生医学替代组织制备领域的需求日益增长。传统子宫内膜上皮细胞与基质细胞的二维单层培养体系，无法复现组织的复杂三维结构。一款可模拟人类子宫内膜微环境的全合成支架，最终可为组织生理学研究提供可靠平台，进而为攻克女性不孕症与体外受精（IVF）失败迈出重要步伐。本研究以高内相乳液模板多孔聚合物（polyHIPEs）作为支架，用于培养原代人子宫内膜上皮细胞（HEECs）与原代人子宫内膜基质细胞（HESCs）。通过组织学实验评估了HEECs与HESCs在细胞接种型polyHIPE支架中的浸润情况，并通过免疫染色验证了细胞表型。共聚焦显微镜观察显示，HEECs与HESCs的形态特征与体内原生状态相符。采用RNA测序分析了polyHIPE支架与单层培养体系中细胞的转录组差异。通过检测子宫内膜标志物基因的表达水平，进一步评估了polyHIPE支架与单层培养体系中HEECs和HESCs的分化状态。本研究结果表明，通过定制化polyHIPE支架，可构建出可模拟天然人类子宫内膜结构与功能的三维细胞培养模型。