Uncovering the Effect of Passage Number on HT29 Cell Line Based on the Cell Metabolomic Approach

Abstract

The passage number is an important factor when designing the cell line-based experiment. Although HT29 cells were widely used in the laboratory for colorectal cancer studies, the impact of cell passage number on the HT29 cells was still unknown. In this study, phenotypic assay and metabolomic approach were applied to analyze the systemic effects of passage numbers (passage 4, 10, and 16) on the HT29 cells. The results showed that the increased cell passage number affected the cell cycle distribution and also decreased the proliferation and migration ability of HT29 cells. The metabolomic analysis coupled with heatmap and hierarchical cluster analysis showed obvious metabolome difference among the cells with different passage numbers, which was related with 61 differential metabolites. Three metabolic pathways were determined as the key pathways, and arginine participated in two of them. In addition, it was found that arginine supplementation could inhibit the proliferation ability of HT29 cells in vitro, and a synergistic effect existed between arginine and cisplatin. In conclusion, this study not only revealed the influence of passage numbers on the HT29 cell but also provided an important reference that arginine has the potential role to be developed as the cisplatin therapeutic adjuvant.

Passage Number of 4T1 Cells Influences the Development of Tumour and the Progression of Metastasis in 4T1 Orthotopic Mice

“…This was in agreement with the study reported by Welch, Neri and Nicolson ( 21 ), whereby it was stated that the metastatic potential of each tumour cell clone drifts with passage number. In 2021, a study by Cao et al ( 22 ) highlighted the influence of increased cell passage number on the proliferation and migration capacity of HT29 cells.…”

Passage number of cancer cell lines: Importance, intricacies, and way‐forward

“…Cao et al (2021) conducted a study using HT29 colorectal cancer cells, revealing a negative correlation between HT29 proliferation and an increasing PN. As the PN increased, there was an observed rise in the population of HT29 cells in the G0/G1 phase, while the cell population in the S and G2 + M phases decreased.…”

Aging Cell Culture - Genetic and Metabolic Effects of Passage Number on Zebrafish Z3 Cells

“…hES were even affected by different passaging methods, such as enzymatic versus nonenzymatic dissociation of cell clusters, the latter of which appeared to induce chromosomal aberrations [11]. Increased passage numbers of HT29 cells were reported to induce a decrease in cell proliferation and their ability to migrate, and led to altered cell cycle distributions [12]. Hence, putative differences in passage numbers that are probably cell-line-specific have to be excluded before starting new experiments.…”