Detection of contaminants in cell cultures, sera and trypsin
Abstract
The aim of this study was standardization and application of polymerase chain reaction (PCR) for the detection of contaminants in cell cultures, sera and trypsin. Five PCR protocols were standardized to assess the presence of genetic material from mycoplasma, porcine circovirus 1 (PCV1), bovine leukemia virus (BLV) or bovine viral diarrhea virus (BVDV) in cell culture samples. PCR reactions for the genes GAPDH and beta-actin were used to evaluate the efficiency of nucleic acid extraction. The PCR protocols were applied to 88 cell culture samples from eight laboratories. The tests were also used to assess potential contamination in 10 trypsin samples and 13 fetal calf serum samples from different lots from five of the laboratories. The results showed the occurrence of the following as DNA cell culture contaminants: 34.1% for mycoplasma, 35.2% for PCV1, 23.9% for BVDV RNA and 2.3% for BLV. In fetal calf sera and trypsin samples BVDV RNA and PCV1 DNA was detected. The results demonstrated that cell culture, sera and trypsin used by different laboratories show a high rate of contaminants. The results highlight the need for monitoring cell cultures and controlling for biological contaminants in laboratories and cell banks working with these materials.
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Animal-derived products may raise contamination issues, for example, due to the presence of viruses.
“Furthermore, animal‐derived products may also raise contamination issues, for example, due to the presence of viruses [20, 21].”
From A worldwide survey on the use of animal‐derived materials and reagents in scientific experimentation (2022) · cites this paper as [21]
Cites
Detection of mycoplasma contamination in cell cultures by a mycoplasma group-specific PCR
van Kuppeveld, F. J., Johansson, K. E., Galama, J. M., Kissing, J., Bölske, G., van der Logt, J. T., & Melchers, W. J. (1994) · Applied and Environmental Microbiology · doi:10.1128/aem.60.1.149-152.1994 · Cited by 186 · see citing works
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Pamies, D. (2018) · ALTEX · doi:10.14573/altex.1710081 · Cited by 127 · see citing works
Jedrzejczak-Silicka, M. (2017) · New Insights into Cell Culture Technology · doi:10.5772/66905 · Cited by 87 · see citing works
A worldwide survey on the use of animal‐derived materials and reagents in scientific experimentation
Cassotta, M., Bartnicka, J. J., Pistollato, F., Parvatam, S., Weber, T., D'Alessandro, V., Bastos, L. F., & Coecke, S. (2022) · Engineering in Life Sciences · doi:10.1002/elsc.202100167 · Cited by 33 · see citing works
- Authors
- Pinheiro de Oliveira, T. F., Fonseca, A. A., Camargos, M. F., de Oliveira, A. M., Pinto Cottorello, A. C., Souza, A. D. R., de Almeida, I. G., & Heinemann, M. B.
- Venue
- Biologicals
- Publisher
- Elsevier BV
- Published
- 2013-11
- Volume
- 41
- Issue
- 6
- Pages
- 407-414
- ISSN
- 1045-1056
- Cited by
- 63
- References
- 1
- Language
- en
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