Mitochondrial Morphology and TOMM20/TIMM44 Abundance During Prolonged HEK293 Cell Culture Under Different Glucose and Medium-Replacement Conditions
- 1Department of Functional Morphology, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo 204-8588, Japan
Abstract
Glucose availability is essential for cellular metabolism and maintaining mitochondrial homeostasis. However, the effects of sustained changes in glucose availability on mitochondrial protein abundance and morphology remain unclear. In this study, we examined the mitochondrial responses to different glucose concentrations and medium replacement conditions. HEK293 cells were cultured for 4 days in standard or high-glucose medium without medium replacement. TOMM20 and TIMM44 abundance was evaluated using immunofluorescence and Western blotting, and mitochondrial morphology was assessed using transmission electron microscopy. The mitochondrial cross-sectional area and density were quantified. Glucose concentrations and medium pH were monitored throughout the culture period. In a separate experiment, the culture medium was replaced every 24 h. Without medium replacement, high-glucose culture was associated with greater TOMM20 and TIMM44 abundance and larger mitochondria than standard-glucose culture, whereas mitochondrial density did not differ significantly. With daily medium replacement, differences in TOMM20 and TIMM44 abundance were no longer detected, and mitochondrial size did not differ significantly between the glucose conditions. Moreover, mitochondrial density did not differ significantly between the glucose conditions. These findings indicate that mitochondrial morphology and TOMM20/TIMM44 abundance during prolonged culture are associated with both the initial glucose concentration and subsequent changes in the extracellular culture environment.
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BibTeX
@article{kawakami2026mitochondrial,
title = {{Mitochondrial Morphology and TOMM20/TIMM44 Abundance During Prolonged HEK293 Cell Culture Under Different Glucose and Medium-Replacement Conditions}},
author = {Kiyoharu Kawakami and Mirai Koike and Nozomi Ito and Rei Watanabe and Takahiro Tsukimura and Kazuhiko Nakadate},
journal = {Cells},
year = {2026},
month = nov,
doi = {10.3390/cells15212031},
url = {https://www.mdpi.com/2073-4409/15/21/2031},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Mitochondrial Morphology and TOMM20/TIMM44 Abundance During Prolonged HEK293 Cell Culture Under Different Glucose and Medium-Replacement Conditions
AU - Kiyoharu Kawakami
AU - Mirai Koike
AU - Nozomi Ito
AU - Rei Watanabe
AU - Takahiro Tsukimura
AU - Kazuhiko Nakadate
PY - 2026
DA - 2026/11/07
JO - Cells
DO - 10.3390/cells15212031
UR - https://www.mdpi.com/2073-4409/15/21/2031
AB - Glucose availability is essential for cellular metabolism and maintaining mitochondrial homeostasis. However, the effects of sustained changes in glucose availability on mitochondrial protein abundance and morphology remain unclear. In this study, we examined the mitochondrial responses to different glucose concentrations and medium replacement conditions. HEK293 cells were cultured for 4 days in standard or high-glucose medium without medium replacement. TOMM20 and TIMM44 abundance was evaluated using immunofluorescence and Western blotting, and mitochondrial morphology was assessed using transmission electron microscopy. The mitochondrial cross-sectional area and density were quantified. Glucose concentrations and medium pH were monitored throughout the culture period. In a separate experiment, the culture medium was replaced every 24 h. Without medium replacement, high-glucose culture was associated with greater TOMM20 and TIMM44 abundance and larger mitochondria than standard-glucose culture, whereas mitochondrial density did not differ significantly. With daily medium replacement, differences in TOMM20 and TIMM44 abundance were no longer detected, and mitochondrial size did not differ significantly between the glucose conditions. Moreover, mitochondrial density did not differ significantly between the glucose conditions. These findings indicate that mitochondrial morphology and TOMM20/TIMM44 abundance during prolonged culture are associated with both the initial glucose concentration and subsequent changes in the extracellular culture environment.
SN - 2073-4409
ER -