normal human liver cells wrl68  (AcceGen Biotechnology)

Journal: Advanced Science

Article Title: A Single‐Cell Metabolic Profiling Characterizes Human Aging via SlipChip‐SERS

doi: 10.1002/advs.202406668

Figure Lengend Snippet: Spermine promotes cellular senescence in vitro. a) Quantification of the nuclear size of GM00038 cells treated with spermine for 14 days. Number of cells per group > 1500, cells from three biological replicates. b) The viability of GM00038 cells was examined by CCK8 after being treated with spermine (1 µg mL −1 ). n = 3 biological replicates. c) Cell cycle analysis of GM00038 cells treated with spermine (1 µg mL −1 ) for 7 days. d) mRNA levels of P16, P21, LMNB1, IL‐6, IL‐8, and IL‐1A in GM00038 cells treated with spermine (1 µg mL −1 ), normalized to GAPDH mRNA (n = 3 per group). e) Representative images of SA‐β‐gal (n = 3 per group/4 images per n), EdU (yellow; n = 3 per group/4 images per n), and DAPI labeled nuclei (blue) of HUVECs treated with spermine (2 µg mL −1 ) for 5 days. Scale bar, 100 µm. f,g) Quantification of SA‐β‐gal + cells (f) and EdU + cells (g) in (d). At least 100 cells per n were calculated. h) mRNA levels of P16, P21, LMNB1, IL‐6, IL‐8, and IL‐1A in HUVECs treated with spermine (2 µg mL −1 ), normalized to GAPDH mRNA. (n = 3 per group). i) Representative images of SA‐β‐gal (n = 3 per group/4 images per n), EdU (yellow; n = 3 per group/4 images per n), and DAPI labeled nuclei (blue) of WRL68 treated with spermine (2 µg mL −1 ) for 12 days. Scale bar, 100 µm. j,k) Quantification of SA‐β‐gal + cells (j) and EdU + cells (k) in (h). At least 800 cells per n were calculated. l) mRNA levels of P16, P21, LMNB1, IL‐6, IL‐8, and IL‐1A in WRL68 treated with spermine (2 µg mL −1 ) for 7 days, normalized to GAPDH mRNA. (n = 3 per group). m) H 2 O 2 levels in GM00038 treated with spermine (1 µg mL −1 ) for 14 days (left, n = 3 per group). The amount of H 2 O 2 released into the cell culture medium within 24 h after GM00038 cells were treated with spermine (1 µg mL −1 ) for 13 days (right, n = 3 per group). n) mRNA levels of SMS, SMOX and SSAT in GM00038 cells treated with spermine (1 µg mL −1 ) for 14 days, normalized to GAPDH mRNA (n = 5 per group). o) Exogenous Spermine treatment could induce typical cellular senescence phenotype and lead to increased overall H 2 O 2 level in vitro. Data are means ± SD of biologically independent samples. Statistical significance was calculated using a two‐tailed t ‐test (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

Article Snippet: Human normal liver cells (WRL‐68) were obtained from ATCC (United States).

Techniques: In Vitro, Cell Cycle Assay, Labeling, Cell Culture, Two Tailed Test