Начало / Университетът / Факултети / Биологически факултет / Официални издания / Годишник на Софийския университет - КНИГА 4 Научни сесии на Биологическия факултет / Том 102, 2017 г. - Младежка научна конференция "Климентови дни" - 2016 г. / Rumiana Ganeva, Dimitar Parvanov, Georgi Stamenov, Balik Dzhambazov, Todor Chaushev - TNF-α EXPRESSION OF IN-VITRO CULTURED HUMAN ENDOMETRIAL STROMAL AND EPITHELIAL CELLS

   

TNF-α EXPRESSION OF IN-VITRO CULTURED HUMAN ENDOMETRIAL STROMAL AND EPITHELIAL CELLS

 

RUMIANA GANEVA1*, DIMITAR PARVANOV1, GEORGI STAMENOV1, BALIK DZHAMBAZOV2,
TODOR CHAUSHEV1

 

1 – Nadezhda, Women’s Health Hospital, 3 “Blaga vest” Street, Sofia, Bulgaria

2 – Department of Developmental Biology, Plovdiv University “Paisii Hilendarski”, Plovdiv, Bulgaria

*Corresponding author:rumqnaganeva1@abv.bg

 

Keywords: TNF-alfa, 3D culture, human endometrial cells, implantation window

 

Abstract: The human endometrium is a complex tissue composed of various cell types, whose interaction determine its main functions in relation to embryo adhesion and implantation. During each menstrual cycle, the human endometrium undergoes a series of well controlled changes in anticipation to the arrival of the blastocyst that are still not completely understood. Previous analyses have shown the presence of TNF-α and its receptors within both the stromal and epithelial cells of the human endometrium. TNF-alpha has a specific menstrual cycle-dependent expression, which is the highest during the secretory phase. This cytokine has a local role in a variety of endometrial functions in order to support embryo implantation, including cell differentiation and tissue remodelling.

In the present study, we examined the secretion of TNF-α by primary human endometrial stromal (ESC) and epithelial (EEC) cells cultured in (1) RPMI1640 media, supplemented with 10% FBS only and (2) RPMI1640 media, supplemented with 10% FBS, progesterone (P4) (1 μmol/L) and 17β-estradiol (E2) (30 nmol/L). Endometrial epithelial and stromal cells were obtained from mid-secretory stage endometrial samples from 28 healthy women and cultured in vitro in 3D Matrigel for 48 hours. The presence of TNF-α in the culture media was measured using enzyme-linked immunosorbent assay (ELISA).

Cell culture measurements revealed that epithelial cells secrete higher quantities of TNF-α (2.2 pg/ml ± 0.7 pg/ml), compared to stromal cells (0.5 pg/ml ± 0.2 pg/ml). Moreover, co-culture of epithelial and stromal cells showed the highest expression of TNF-alpha (16.5 pg/ml ± 3.2 pg/ml). Additionally, P4 and E2 supplementation raised the TNF-alpha level in the culture media.

These findings suggest that co-culture of these primary cell types has a synergetic effect on the enhancement of TNF-α secretion. Its higher expression in the co-culture proves that the combination of both cell types is far more relevant to in vivo conditions, compared to primary ESC or EEC lines cultured separately. Furthermore, culturing ESC and EEC together in the context of hormone presence and 3D culture conditions establishes an adequate architectural and functional in vitro model for studying of certain aspects of endometrial receptivity and implantation in humans.

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