Abstract

COBAXIN CARDENAS, Mayra Elizeth et al. In vitro doubling time determination of BUVEC-E ₆ E ₇ cell line of bovine origin. Rev. mex. de cienc. pecuarias [online]. 2018, vol.9, n.1, pp.162-170. ISSN 2448-6698.  https://doi.org/10.22319/rmcp.v9i1.4316.

It has been suggested that rickettsia Anaplasma marginale might interact with endothelial cells in the bovine host. BUVEC E6E7 endothelial cell line has been utilized to grow Anaplasma marginale with no success, partially because in vitro growth characteristics were unknown. Therefore, several experiments were assayed in an attempt to retrieve useful information to define them. Growing culture cells were obtained from UNAM’s Neurobiology Institute, maintained with fetal calf serum and NaHCO3 supplemented F-12 (HAM) medium and incubated at 37 °C on 5% CO2-air humid mixture atmosphere. First two assays used 24 well plates were focused on determination of a minimum initial cell concentration, which was 2,500 cells/well and cell density, whose experiment was set with 1, 3, 9 and 27 cells/mm2. Cells were harvested with an EDTA-Trypsin solution when confluence was reached. Average final concentration of 8.0 x 104, 9.3 x 104, 6.3 x 104 & 6.5 x 104 cells/well was respectively found. 35 mm Ø sterile Petri dishes were used for kinetics studies. Cultures were set in triplicate with a seed density 3 cell/mm2 divided into two series and sampling every 8 h. A 48 h culture media replacement interval applied for the first series and none for the second. Maximum growth was observed in both series at 152 h with 11.73 and 12.6 h doubling time in culture, respectively. It was confirmed that BUVEC E6E7 endothelial cell line expresses unusual growth strength.

Keywords : Doubling time; BUVEC-E₆E₇; In vitro culture.