Cultures of bovine endometrial cells and macrophages for investigating cellular interactions

Description

This study aimed to establish a primary bovine endometrial cells culture and a monocyte-derived macrophages culture followed of a co-culture system of both cellular types. Our approach provides a simple protocol, cost-effective, and require no specialized equipment. Therefore, these cultures developed are a useful in vitro model for investigating the cellular interactions and modulating cell endometrial functions.

Steps to reproduce

The May Grumwald Giemsa staining involved cells fixation, May-Grünwald and Giemsa solution, and microscopy. Immunocytochemistry used primary antibody specific for cytokeratin (epithelial) or vimentin (stromal) cells. Also, Flow Cytometry was used to confirm the identity of endometria cells in culture. The cells were resuspended in focusing buffer and acquired using flow cytometer. The gating strategy was used to analysis. Mononuclear cells isolated from peripheral blood of cows were seeded. Centrifugation facilitated adherence of monocytes to the plate. Non-adherent cells were removed. Presumed macrophages was continued for 6 days.

Categories

Funders

• Agencia Nacional de Promoción de la Investigación, el Desarrollo Tecnológico y la Innovación

  Argentina

  Grant ID: PICT 2020-3403

Licence