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• W2000462040 abstract "Objective: Evaluation of biodegradable three-dimensional (3-D) matrixes for tissue engineering of human endometrium. Design: Endometrial cells (2 × 105) were seeded in a highly porous extracellular matrix composed of type I collagen and chondrotin-6-sulfate (CG copolymer matrix) and cultured for 1–3 weeks. The cellular growth pattern was analyzed by histology, propidium iodide (PI) labeling and immunostaining with the antibodies of desmin (specific for stromal cells) and pan-cytokeratin (specific for glandular epithelial cells). Image Pro plus software was used for data analysis. Materials/Methods: Enzymatic isolated human endometrial stroma and gland cells (1:1 ratio) were seeded on CG copolymer matrixes and cultured in 24 well microtiter plates for 2 weeks in RPMI 1640, Ham’s F10, or DMEM with 10% fetal bovine sera ± insulin (IU/ml). Next, these cells were also cultured in media containing DMEM + insulin, plus estradiol (E2) (200 pg/ml) or E2 + 6α-methyl-17α-hydroxy-progesterone acetate (MPA) (0.2 uM) for 1 to 3 weeks. After culture, the matrixes were fixed with 10% neutral buffered formalin, embedded in paraffin and sectioned at 5μ. Every sixth slide was labeled with PI to examine the total area of cell growth utilizing the Zeiss fluorescent axioskop and the Image Pro plus program. The slides adjacent to that of PI staining were used for pan-cytokeratin and desmin immunoassay to assess the population of glands and stromal cells during culture in-vitro. Results: The seeding cells were found to be capable of attaching to and surviving within the porous copolymer matrix to form a full thickness of three-dimensional tissue. After 2 weeks of culture, total cell area (Pixel × 106/0.5 mm2 matrix) in culture with or without insulin were 0.9 and 2.0 in DMEM; 0.4 and 1.5 in RPMI 1640; and 0.5 and 0.7 in Ham’s F10, respectively. During 1 to 3 weeks of culture in DMEM plus insulin, the total cell area increased from 0.7 to 2.5 with no hormone supplementation from 0.7 to 2.9 with supplementation of E2, and from 0.5 to 4.4 with supplementation of E2 + MPA, respectively. Fluorescent dual-staining further reveals that the population of pan-cytokeratin positive cells (i.e., gland cells) was greatly enhanced by steroids. After 3 weeks culture in DMEM + Insulin, the population of glands in the 3-D structures were 20 ± 4% with no hormone, 34 ± 2% with E2, and 57 ± 2% with E2 + MPA supplementation. Conclusions: We have successfully engineered human endometrial tissue using a biodegradable polymer as a cell carrier. Insulin and steroids appear to be essential for 3-D endometrial cell growth in-vitro. With exogenous steroids, these cells were able to reorganize in the porous extracellular matrix mimicking in-vivo endometrium to form glandular-like structures. We can foresee that this three-dimensional endometrial culture can serve as an in-vitro model to study the mechanism of implantation and interaction between the embryo and endometrium." @default.
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• W2000462040 date "2001-09-01" @default.
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• W2000462040 title "Tissue engineering of human endometrial cells using a biodegradable polymer." @default.
• W2000462040 doi "https://doi.org/10.1016/s0015-0282(01)02091-x" @default.
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