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.