Department of Materials Science and Engineering , Monash University , Clayton , 3800 , Victoria Australia.
Department of Polymers, Chemical Industries Research Division , National Research Centre (NRC) , El Bohouth St. 33 , Dokki, Giza, 12622 , Cairo , Egypt.
Biomacromolecules. 2018 Aug 13;19(8):3343-3350. doi: 10.1021/acs.biomac.8b00635. Epub 2018 Jul 10.
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.
需要开发新方法来体外培养原代人细胞，以便研究细胞和组织生理学，并为再生医学培养替代组织。常规的二维单层培养子宫内膜上皮细胞和基质细胞无法复制组织的复杂 3D 结构。模仿人子宫内膜微环境的全合成支架最终可以为研究组织生理学提供强大的平台，从而为解决女性不孕和试管婴儿失败问题迈出重要一步。在这项工作中，我们研究了乳液模板化多孔聚合物（称为聚 HIPEs）作为原代人子宫内膜上皮细胞和基质细胞（HEECs 和 HESCs）培养的支架。通过组织学研究评估 HEECs 和 HESCs 渗透到细胞接种的聚 HIPEs 支架中的情况，并通过免疫染色确认表型。共聚焦显微镜显示，HEECs 和 HESCs 的形态与体内观察到的形态相似。通过 RNA 测序研究了在聚 HIPEs 支架上和单层培养物中生长的细胞之间的转录组差异。通过监测子宫内膜标记基因的表达，进一步评估了在聚 HIPEs 支架和单层培养物中生长的 HEECs 和 HESCs 的分化状态。我们的观察结果表明，可以使用定制的聚 HIPEs 支架开发出一种能够模拟天然人子宫内膜结构和功能的 3D 细胞培养模型。
