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It has been suggested that rickettsia Anaplasma marginale might interact with endothelial cells in the bovine host.  BUVEC E 6 E 7 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 NaHCO 3 supplemented F-12 (HAM) medium and incubated at 37 °C on 5% CO 2 -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/mm 2 .  Cells were harvested with an EDTA-Trypsin solution when confluence was reached. Average final concentration of 8.0 x 10 4 , 9.3 x 10 4 , 6.3 x 10 4 & 6.5 x 10 4 cells/well was respectively found. 35 mm O sterile Petri dishes were used for kinetics studies. Cultures were set in triplicate with a seed density 3 cell/mm 2 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 E 6 E 7 endothelial cell line expresses unusual growth strength.

Determinación del tiempo de duplicación in vitro de la línea celular BUVEC-E6E7 de origen bovino