Dr. Alshammari states for the experimental basis of his study: “Chang and Huh-7 liver cell lines were utilized as human normal hepatocyte model and innovation to mimic the liver environment” [1]. Based on his findings he concludes “The findings of the current study are critical for the development of novel herbal-based therapies for fatal disorders such as liver cancer” [1].

Although the data shown is certainly not wrong, one critical needs attention. The Chang liver cell line used in this study was reported to be derived from normal liver [2]. However, the first doubts about the origin of this cell line were already published in 1967 [3]. The typical short tandem repeat profile and other genetic characteristics have later shown that this cell line is a de facto strain of HeLa [4,5]. Authors working in the field of Hepatology have realized this fact and the Register of Misidentified Cell lines lists the Chang liver line in its register as a misidentified cell line under register number ICLAC-00002 [7,8]. Therefore, it is somewhat surprising that Alshammari used this cell line as a “normal innovative hepatocyte model”.

Consequently, the conclusion that these findings are specific for hepatocellular carcinoma cells is highly questionable. Quite the opposite is correct because the cytotoxic effects of the seed extracts of L. angustifolia were the quite same in hepatoma cell line HuH-7 and in the HeLa derivative “Chang liver”.

Unfortunately, cell line misidentification is a relentless problem in the field of biomedical research provoking potentially false conclusions and irreproducible results not only in this study [8]. It is estimated that about one fifth to one third of all used cell lines are misidentified [8]. This terrifying shortcoming can only be remedied by raising the awareness of authors and reviewers to this issue.

References cited
1. Alshammari GM. Cytotoxic effects of Lavandula angustifolia seed extracts on the viability of Huh-7 and Chang liver cells. PLoS One 2022;17(4):e0267499. doi: 10.1371/journal.pone.0267499
2. Chang RS. Continuous subcultivation of epithelial-like cells from normal human tissues. Proc Soc Exp Biol Med. 1954;87(2):440-3. doi: 10.3181/00379727-87-21406
3. Gartler SM. Genetic markers as tracers in cell culture. Natl Cancer Inst Monogr. 1967;26:167-95
4. Nelson-Rees WA, Flandermeyer RR. HeLa cultures defined. Science 1976;191(4222):96-8. doi: 10.1126/science.1246601
5. Masters JR, Thomson JA, Daly-Burns B, Reid YA, Dirks WG, Packer P, Toji LH, Ohno T, Tanabe H, Arlett CF, Kelland LR, Harrison M, Virmani A, Ward TH, Ayres KL, Debenham PG. Short tandem repeat profiling provides an international reference standard for human cell lines. Proc Natl Acad Sci U S A 2001;98(14):8012-7. doi: 10.1073/pnas.121616198
6. Gao Q, Wang XY, Zhou J, Fan J. Cell line misidentification: the case of the Chang liver cell line. Hepatology 2011;54(5):1894-5. doi: 10.1002/hep.24475
7. ICLAC. Register of Misidentified Cell lines. Available at: https://iclac.org/databas... (last accessed 20 July 2022)
8. Horbach SPJM, Halffman W. The ghosts of HeLa: How cell line misidentification contaminates the scientific literature. PLoS One 2017;12(10):e0186281. doi: 10.1371/journal.pone.0186281

Ralf Weiskirchen
RWTH University Hospital Aachen
Institute of Molecular Pathobiochemistry, Experimental Gene Therapy and Clinical Chemistry (IFMPEGKC)
Aachen, Germany
Email: rweiskirchen@ukaachen.de
Phone: +49-(0)241-8088683