Calcium imaging in C2C12 cells and RAW264.7 cells post co-culture and changes induced by sodium palmitate
10.3969/j.issn.1672-8467.2025.06.013
- VernacularTitle:C2C12细胞和RAW264.7细胞共培养后细胞内钙成像及棕榈酸钠诱导后的改变
- Author:
Li-jun SONG
1
;
Shuang WU
;
Qin SHA
;
Chuan-xin YANG
;
Xing-yu TONG
;
Hui JIANG
Author Information
1. 复旦大学附属中山医院青浦分院麻醉科 上海 201700
- Publication Type:Journal Article
- Keywords:
C2C12 cell;
RAW264.7 cell;
co-culture;
calcium imaging;
sodium palmitate(PA);
Paraxanthine
- From:
Fudan University Journal of Medical Sciences
2025;52(6):877-882
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the effect of RAW264.7 cells on calcium sparks in a insulin resistance model of C2C12 cells induced by sodium palmitate.Methods C2C12 cells and RAW264.7 cells were co-cultured to simulate the in vivo state of skeletal muscle.C2C12 cells were cultured in high-glucose medium containing 2%horse serum to induce differentiation into mature myotubes,and then divided into 5 groups:control(RAW264.7 cells),co-culture of C2C12 with RAW264.7,C2C12 alone,co-culture of C2C12 with RAW264.7 plus sodium palmitate(PA),and C2C12 alone with PA.PA of 5 mmol/L was used to induce insulin resistance in C2C12 cells for 24 hours.Revived and expanded RAW264.7 cells were evenly added to C2C12 cells and co-cultured for two days.Subsequently,cells were maintained in modified suspension culture,and both cell types were loaded with the calcium ion fluorescent probe Fluo-4 AM.Finally,Paraxanthine was used to induce intracellular calcium sparks,which was captured and recorded under a laser confocal microscope.Results No significant calcium signal change was observed in the control group.Co-cultured C2C12 cells exhibited rapid and pronounced calcium signal changes,whereas calcium signals in C2C12 cells cultured alone increased slowly throughout the observation period without a sharp decline.The peak calcium signal was reached significantly faster in co-cultured C2C12 cells than that in C2C12 cells cultured alone(P<0.001).With PA induction,calcium signal changes in C2C12 cells were not markedly altered,while distinct calcium fluctuations were still observed in co-cultured C2C12 cells,and the peak calcium signal was reached significantly faster in co-cultured C2C12 cells than that in C2C12 cells cultured alone(P<0.001).Conclusion RAW264.7 cells enhance the dynamic responsiveness of calcium signaling in both normal and PA-stimulated C2C12 cells.
