Fig. 5
Endometrial three-dimensional cell culture model. a Microscopic appearance of endometrial stromal cells embedded in atelocollagen on Day 1. Cell numbers were 1 × 106cells (left) and 2 × 106cells (right), respectively. Black bar is 500 μm. b Gross appearance of endometrial stromal cells embedded in atelocollagen on day 7. Cell numbers were 1 × 106cells (left) and 2 × 106cells (right), respectively. Black bar is 1 cm. c Microscopic appearance (bright field and HE staining) of endometrial stromal cells embedded in atelocollagen on day 7. Cell numbers were 1 × 106cells (left) and 2 × 106cells (right), respectively. Black bar is 1 cm. d Protocol for three-dimensional cell culture. e Gross appearance of endometrial three-dimensional cell culture. Endometrial stromal cells were embedded in atelocollagen (left), then endometrial epithelial cells were plated on formed stromal layers using a glass ring on day 7 (middle). Endometrial three-dimensional model was developed during further 14 days of culture (right). f HE staining for three-dimensional cultured endometrial cells. Black bar is 100 μm. g Magnification of box area in figure f. Endometrial epithelial cells (arrows) and stromal cells (arrowheads) in three-dimensional cell culture. Black bar is 50 μm. h Immunohistochemistry of endometrial cells in three-dimensional culture. The endometrial epithelial cells were positive for pan-cytokeratin, vimentin, E-cadherin, and CD13. This is consistent with expression of these markers in intact endometrial tissue. Nuclei were stained with DAPI. Yellow bar is 200 μm