Enhanced Differentiation Potential of Primary Human Endometrial Cells Cultured on 3D Scaffolds
Enhanced Differentiation Potential of Primary Human Endometrial Cells Cultured on 3D Scaffolds
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DOI：
10.1021/acs.biomac.8b00635
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发表时间：
2018-08-01
影响因子：
6.2
通讯作者：
Cameron, Neil R.
中科院分区：
文献类型：
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作者：
Eissa, Ahmed M.;Barros, Flavio S. V.;Cameron, Neil R.
越来越需要体外培养原代人类细胞的新方法来研究细胞和组织生理学，并为再生医学生长替代组织。传统的子宫内膜上皮细胞和基质细胞的2D单层培养无法复制组织的复杂3D结构。模拟人类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一个强大的平台，从而为解决女性不孕症和IVF失败采取重要措施。在这项工作中，乳液模板化的多孔聚合物（称为polyHlPE）被研究作为用于培养原代人子宫内膜上皮细胞和基质细胞（HEEC和HESC）的支架。通过组织学研究评估HEEC和HESC向细胞接种的polyHIPE支架中的浸润，并通过免疫染色确认表型。共聚焦显微镜显示HEEC和HESC的形态是体内发现的代表。使用RNA测序来研究在polyHIPE支架上生长的细胞和在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志物基因的表达，进一步评估在polyHIPE支架和单层培养物中生长的HEEC和HESC的分化状态。我们的观察结果表明，可以使用定制的polyHIPE支架开发一个3D细胞培养模型，可以近似天然人子宫内膜的结构和功能。
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 polyHlPEs) 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.