We, the RIKEN BioResource Center, regret to inform you that 23CLN cell line which had been distributed as a bovine cell line is not the bovine origin.
23CLN was deposited as a bovine cell line in 1987 and had been distributed since then. Immediately after the deposition, we examined the species origin of 23CLN by the conventional isozyme analysis of two different isozymes, lactate dehydrogenase and nucleotide phosphorylase, similarly to other cell banks such as ATCC in USA. The results were interpreted that 23CLN was the bovine origin.
The isozyme analysis depends on the biochemical feature of certain enzymes that are common to several animal species, but show different pattern in electrophoresis. Recently, a more robust molecular method, the species-specific PCR analysis of mitochondria DNA, was established to identify the origin of animal species of cell lines (Ref. 1). We have used this molecular method to test and confirm the origin of deposited animal cell lines since 2011. However, for the cell lines deposited and tested before 2011, we have relied solely on the results of the isozyme analysis and never tested the molecular method.
Very recently, a user of 23CLN informed us that one could not amplify bovine genes from DNA of 23CLN by PCR using specific primers. To confirm the information, we performed the species-specific mitochondria DNA analysis of 23CLN. The results showed that 23CLN was derived from porcine but not from bovine. We also tested the cells cryopreserved immediately after the deposition, and the results were same indicating that this misidentification occurred prior to the deposition in our center.
We have sent notice to all past users of 23CLN by March 4, 2015, expressing our sincere apology and explaining this matter.
Based on this incident, we decided to apply the species-specific mitochondria DNA analysis to animal cell lines that have been deposited before 2011, except for those of human and mouse origin. The test will be performed prior to the provision whenever applicable.
For human cell lines, we introduced short tandem repeat (STR) polymorphism analysis in 2004 (Ref. 2). For mouse cell lines derived from inbred strains, we developed and introduced simple sequence length polymorphism (SSLP) analysis to identify the originated strain of each cell line (Ref. 3).
If you have any question and concern on this matter, please feel free to contact us (yukionakbrc.riken.jp).
- Ono, K. et al. Species identification of animal cells by nested PCR targeted to mitochondrial DNA. In Vitro Cell Dev Biol Anim. 43: 168-175 (2007)
- Yoshino, K., et al. Essential role for gene profiling analysis in the authentication of human cell lines. Hum. Cell 19: 43-48 (2006)
- Yoshino, K., et al. Development of a simple method to determine the mouse strain from which cultured cell lines originated. Interdisciplinary Bio Central 2: Article No. 14 (open access journal) doi: 10.4051/ibc.2010.2.4.0014 (2010)