The necessity of identity assessment of animal intestinal cell lines: A case report

Abstract

Eight intestinal cell lines, established from different animal species were submitted to DSMZ (German Collection of Microorganisms and Cell Cultures) in order to analyze their species of origin and their microbial contamination. Species identity was determined by PCR targeting mitochondrial genes and hence confirmed by sequencing the amplified PCR products. For three cell lines (CIEB, CLAB, PSI-1) we confirmed the species identity, whereas the species of origin of the three other cell lines (B6, B10XI and IPEC) was not the expected one: B6 and B10XI cells, which were supposed to be of chicken origin were identified as porcine cells. IPEC, allegedly a sub clone of the well-known porcine intestinal cell line IPEC-J2, was of bovine instead of porcine origin. However, two further IPEC-clones, namely IPEC-1 and IPEC-J2, provided by another source were shown to be derived from the correct species (i.e. pig). Furthermore, six out of these eight cell lines turned out to be highly contaminated with mycoplasma. Alerted by this high incidence of infected and false specified cell lines, we feel obliged to inform all those working with animal intestinal cell lines and we strongly recommend verifying the species identity before using them. Also, the presence of mycoplasma should be tested when taking the cells in culture for the first time, and this mycoplasma control should be repeated at regular time intervals (e.g. every 4 weeks).

Physiological Concentration of Exogenous Lactate Reduces Antimycin A Triggered Oxidative Stress in Intestinal Epithelial Cell Line IPEC-1 and IPEC-J2 In Vitro

“…Next, we analysed functional consequences of such lactate concentrations in neutral pH on enterocytes and utilized intestinal porcine cell lines IPEC-1 and IPEC-J2. Both cell lines are of small intestine origin and can therefore be used to mimic the in vivo situation in pig [ 18 ]. In consequence of the in vivo studies we chose a lactic acid concentration of 25 mmol/L as a realistic level for further in vitro studies.…”

“…Cells were routinely seeded at a density of 0.6 x 10 5 /mL (IPEC-1) with 7.5 mL medium in plastic tissue culture flasks (25 cm 2 Nunc) and passaged every 3–4 d for a maximum of 20 times (IPEC-1 passages 103-123). Additionally, ROS measurements were performed in parallel with IPEC-J2 (ACC701) cells [ 18 ]. IPEC-J2 cell line was cultured as described for IPEC-1 except a lower seeding density of 0.5 x 10 5 /mL.…”

Deoxynivalenol Affects Cell Metabolism and Increases Protein Biosynthesis in Intestinal Porcine Epithelial Cells (IPEC-J2): DON Increases Protein Biosynthesis

“…Intestinal porcine epithelial cells (a friendly gift from Mariana Roselli, Rome, Italy; IPEC-J2 ACC 701; [ 55 , 56 , 57 ]) were regularly tested and found to be free of mycoplasma contamination (Venor GeM Mycoplasma Detection Kit; Minerva Biolabs, Berlin, Germany). DMEM F12 (1:1) (with glutamine: 2.5 mM, with D-glucose: 3.16 g/L) supplemented with 5% fetal bovine serum (FBS), 16 mM 4-(2-hydroxyethyl)-1-piperazineethansulfonic acid (HEPES), 1% insulin-transferrin-selenium (ITS) (all from PAN-Biotech, Aidenbach, Germany), and 5 ng/mL epidermal growth factor (EGF, Biochrome, Berlin, Germany) was used as cell culture medium (glucose concentration: 3.16 g/L; [high]).…”

Comparing Two Intestinal Porcine Epithelial Cell Lines (IPECs): Morphological Differentiation, Function and Metabolism

“…Intestinal porcine epithelial cells (IPEC-1 ACC 701 and IPEC-J2 ACC 705; [ 10 ], Leibniz Institute DSMZ—German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany) were regularly tested and found to be free of mycoplasma contamination (Venor GeM Mycoplasma Detection Kit; Minerva Biolabs, Berlin, Germany). In all experiments, cells were seeded with a density of 0.88*10 5 /cm 2 on permeable support (ThinCerts; pore size: 1 μm; polyester; Greiner bio-one, Germany).…”