Detection of contaminants in cell cultures, sera and trypsin

作者：Pinheiro de Oliveira Tatiana Flavia; Fonseca Antonio Augusto Jr; Camargos Marcelo Fernandes; de Oliveira Aranapolino Macedo; Pinto Cottorello Ana Claudia; Souza Antonizete dos Reis; de Almeida Iassudara Garcia; Heinemann Marcos Bryan^*

来源：Biologicals, 2013, 41(6): 407-414.

DOI：10.1016/j.biologicals.2013.08.005

• Viral contaminants
• Mollicutes
• PCR
• Cell cultures
• Sera
• Trypsin

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 23% 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.

• 出版日期2013-11