Science and Technology Production

Medicina - ESTABLISHING A 3D MOUSE PRIMARY CELL CULTURE MODEL TO STUDY THE DIRECT EFFECTS OF ENVIRONMENTAL ESTROGENS ON THE FUNCTIONAL DIFFERENTIATION OF THE MAMMARY GLAND

Congress

Authorship:

Altamirano GA ; GOMEZ, AYELEN LUCIANA ; Rodriguez HA ; Luque EH ; Muñoz-de-Toro M ; Kass L

Date:

2017

Publishing House and Editing Place:

Sociedad Argentina de Investigación Clínica

Summary *

There are several human-made chemical compounds with hormonalaction. Among them, Bisphenol A (BPA) and benzophenone-3(BP3) have been shown to have estrogenic activities. Previously,we had demonstrated that the development and differentiation ofthe mammary gland are affected by perinatal exposure to xenoestrogensin vivo. In the present study, our aim was to developand establish an in vitro culture model to study the direct effects ofenvironmental estrogens during the functional differentiation of themammary gland. Mammary organoids were obtained and isolatedfrom 8 weeks-old C57BL/6 female mice using enzymatic and mechanicaldigestion. The organoids were cultured on top of Matrigelwith growth medium for 24 hs at 37°C, 5% CO2. The next day, themedium was changed to a differentiation one containing lactogenichormones and exposed to: vehicle (1% ethanol), 1 μM of BPA and1 μM or 1 ηM of BP3 for 72 hs. The mRNA expression of estrogenalpha (ERS1), progesterone (PR) and prolactin receptors (PRLR)and the mRNA levels of the milk protein beta-casein (CSN2) wereanalyzed by real time RT-PCR. Also, CSN2 and alpha-lactoalbumin(LALBA) protein expression was quantified in histological sectionsby an immunofluorescence assay. After 72 hs of exposure with 1μM BPA, ESR1 and PR mRNA levels were like controls, but CSN2mRNA and protein expression was increased, indicating that this3D culture model responded to BPA treatment during differentiation.Furthermore, the exposure to both doses of BP3 induced an increaseof ERS1, PR and PRLR expression; and 1 μM BP3 augmentedCSN2 and 1 ηM BP3 increased LALBA protein expression comparedto controls. In conclusion, our results show that a 3D mouseprimary cell culture model could be an appropriate tool to study thedirect effects of environmental estrogens, and demonstrate that directexposure to low doses of BPA and BP3 induce alterations onmammary gland functional differentiation and milk protein synthesis. Information provided by the agent in SIGEVA

Key Words

MAMMARY GLAND3D CELL CULTUREENVIRONMENTAL ESTROGENSMILK PROTEINS