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Most cell lines have been established from rodents and humans. While DNA fingerprinting may be applied to authenticating human lines, the lack of genetic variation in inbred rodent strains in particular, and our ignorance regarding subspecies differences in non-humans demands different approaches among animal cell lines.

Figure 1: Agarose gel electrophoresis after species PCR with primer pairs targeting canine mitochondrial sequences. An unknown sample (lane 6) was submitted to the DSMZ for species determination. Originally the sample was assumed to be of human origin. However, analysis with human STR probes excluded a human origin, subsequent species PCR indicating a canine origin.

In the past we have developed PCR-based analyses for the discrimination of human, mouse, rat and hamster cell lines. Current research addresses: i) cell lines derived from additional species and; ii) developing methods for distinguishing primate cell lines.

Although cell lines from rodents and humans are those most commonly encountered, it is important to identify other species from which cell lines have been established. Mitochondrial genome sequencing has facilitated this option. We and others have established standard and multiplex-PCR for animal cell lines (as depicted in Figure 1). Now, we are trying to extend the scope of animal cell line species detectible by PCR and to determine the species origin of unknown cell lines submitted to DSMZ (see Table 1).

References:

1. Steube KG, Koelz AL, Drexler HG. Identification and verification of rodent cell lines by polymerase chain reaction. Cytotechnology 56: 49-56 (2008).

2. Steube KG, Meyer C, Uphoff CC, Drexler HG. A simple method using ß-globin polymerase chain reaction for the species identification of animal cell lines - A progress report. In Vitro Cell Dev Biol 39: 468-475 (2003).

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Dr. Steube, Klaus
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Table 1: Animal Species which can be detected by PCR or which are being investigated
Bombyx mori silkworm
Bos taurus bovine
Canis spec. dog
Drosophila melanogaster common fruit fly
Equus caballus horse
Felis spec. cat
Gallus domesticus chicken
Macaca mulatta rhesus monkey
Mesocricetus auratus Syrian hamster
Oncorhynchus mykiss rainbow trout
Ovis aris sheep
Sus scrofa pig
Saguinus oedipus cotton top tamarin
Spodoptera frugipera fall armyworm
Xiphophorus Xiphidium swordtail platy
Xenopus laevis South African clawed toad