Un método alternativo para el aislamiento de Células Luteales Bovinas en alta pureza: centrifugación en gradiente de densidad discontinuo de Percoll (GDDP)

MARURI, A.; BEZENZETE, G.; TEPLITZ, G.; LOMBARDO, DM.

Ciudad Autónoma de Buenos Aires

Congreso; Primer Congreso de la Sociedad Latinoamericana de Reproducción Animal (SOLARA); 2015

Sociedad Latinoamericana de Reproducción Animal

Corpus luteum (CL) is a transient endocrine gland, established by residual follicular wall cells (granulosa and theca cells) following ovulation. The main function of CL is the production of progesterone, a hormone which regulates various reproductive functions. There are at least four distinct morphological cell types: large luteal cells, small luteal cells,capillary endothelial cells, and fibroblast cells. The aim of this study was to evaluate the efficacy of a PDDG, to purify luteal cells. Bovine CL from early luteal phase were treated by mechanical and enzymatic digestion to obtain a primary culture. Once the cells wereconfluent, were trypsinized and centrifuged in a PDDG (10%, 20%, 30%, 35%, 37.5% and 40%). The purified cells, called Passage 1 (P1), were identified by Immunocytochemistry for3?-hydroxysteroid dehydrogenase. We found a 100% purity in Percoll density layers 5 and 6 clearly higher than standard technique without selection with Percoll that only reached 80.6%±7.02 purity. This method offers a cost effective alternative to isolate luteal cells in high-purity to be able to use as in vitro model in various assays.