Abstract
The immortalized cell lines derived from human embryonic kidney, named HEK 293, are extensively used as models of human renal cells in in vitro studies. Nevertheless, ample evidence in the literature shows that HEK 293 cells display genotypic and phenotypic characteristics that differ substantially from primary kidney cells, with potential detrimental effects on the quality of the experimental results. Among the differences documented between HEK 293 and renal cells, there is an altered pattern of expression of many proteins involved in the development and physiological functions of the kidney. Methionine sulfoxide reductase (Msr) enzymes are ubiquitous components of the cellular machinery, evolved to counteract the damages inflicted to methionine residues by oxidative stress, particularly intense in kidney tissues. In this article, we have compared the levels of expression of several different Msr enzymes in human kidney and in a HEK 293 strain and have observed significant differences between the two cell types.
Highlights
In 1951, the first immortalized human cell strain was developed by propagation of poliomyelitis virus in tissue cultures prepared from epidermoid carcinoma of the cervix [1]
We examined the mRNA levels of MsrA, MsrB1, and MsrB2, the typical forms of Methionine sulfoxide reductase (Msr) expressed in mammalian tissues, and for which the presence in the human renal tissues has been documented for adult and embryonic kidney [11,12,13]
The presence of MsrA, MsrB1, and MsrB2 transcripts was evaluated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in cDNA obtained from HEK 293T cells and total human kidney
Summary
In 1951, the first immortalized human cell strain (designed as HeLa) was developed by propagation of poliomyelitis virus in tissue cultures prepared from epidermoid carcinoma of the cervix [1]. Immortalized cell lines are extensively used in many branches of basic biomedical research, from the study of gene function to generation of artificial tissues. They have several advantages over primary cells, such as the ability to proliferate virtually indefinitely, cost-effectiveness, easiness of handling, and no ethical restrictions associated with the use of animal or human tissues. Among the many advantages there are, some critical issues that could dramatically affect the goodness of the results, such as contamination by other cell lines or by mycoplasma, and the misidentification of cell lines About these problems, it is estimated that up to one third of cell lines are imposters [2,3,4,5]
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