Establishment and Characteristics of a Cell Line from Fetal Bovine Thyroid (FBTY)

Abstract

Fischer 0., M. Machatkova, L. Pecuchova, D. Zendulkova and Z. H 0 fin 0 va: Establishment and Characteristics ofa Cell linefrom FetalBovine Thyroid (FBIT). Acta vet. Bmo, 63, 1994: 81-87.Characteristics of a cell line from fetal bovine thyroid were tested in the 150th passage. The line was adapted to growth in the minimal essential medium supplemented with 10 % bovine fetal semm and it had an epithelioid morphology. After seeding of lxlO" cellslml, the culture density was higher than lxl()f> cellslrnl on day 6. The FBTY line was heteroploid. The average number of chromosomes was 48.8 ± 10.0 and only 1 % of mitoses had a diploid number of chromosomes (2n = 60). Species specificity (Bos taurus) was confirmed by indirect immunofluorescence. Besides pronounced fluorescence with rabbit antisera against bovine proteins, weak reactions with antisera against ovine, caprine and swine proteins were observed. The FBTY line was susceptible to two strains of parainfluenza 3 virus, six strains of infectious bovine rhinotracheitis virus and bovine adenoviruses of the serovars I, 2, 3, and 8. • line ECTC derived from fetal bovine thyroid gland which was, in passage 150, susceptible to all known serovars of bovine adenoviruses, including members of the subgroup II which are difficult to replicate. Mitosis, indirect immunofluorescence, bovine adenovirusIn 1981, Marie Machatkova in the Veterinary Research Institute, Bmo, established a cell line from fetal bovine thyroid gland and the results are described in this paper.The characteristics of this cell line (FBTY) were tested in passage 150.Replications of bovine adenoviruses in FBTY and some other bovine cell cultures were compared by P e c uchova (1993). Materials and Methods Establishment and cultureThyroid gland of a nine-month-old male bovine fetus was collected under sterile conditions immediately after exsanguination (Machatkova and PospBil 1975) and transported to the laboratory in minimal essential medium (MEM SEVAC, Institute of Sera and Vaccines, Praha) supplemented with 200 I. U. of penicillin and 200 \lg of streptomycin per m!. Tissue fragments were washed in the medium and cultured in 60 mm plastic Petri dishes in 5 ml of the EPL SEVAC medium (Institute of Sera and Vaccines, Praha) containing 10 % bovine fetal serum, 100 I. U. of penicillin and 100 \lg of streptomycin per m!. The cells were cultured at 37°C in an atmosphere containing 5 % CO 2 , The EPL was replaced after 72h by a 1: 1 mixture of EPL+MEM subsequently changed every other day.As soon as the attached fragments were surrounded by growing cells, the cells were released by 0.02 % versene with 0.25 % trypsin Difco (1:250), reseeded into Mueller's flasks and cultured further in MEM with 10 % bovine fetal serum, 100 I. U. of penicillin and 100 \lg of streptomycin per ml in 300 ml and 500 rnl Roux flasks, Legroux flasks and Mueller's flasks.In passage 85, the cells were frozen and kept for 8 years (1981)(1982)(1983)(1984)(1985)(1986)(1987)(1988)(1989) in liquid nitrogen. When reseeded after passage 85,...