Human Uterine Myocytes Retain Their Energy Charge with No Gross Alterations in Morphology for at Least 8 Days when Cultured under Anaerobic Conditions

Olovsson, Matts; Nordling, Margareta; Ulmsten, Ulf; Lindblom, Bo; Waldenström, Anders; Ronquist, Gunnar

doi:10.1159/000010240

Cited by 4 publications

(1 citation statement)

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“…High concentrations of lactate in culture medium of uterine biopsies incubated for prolonged time intervals (days) were detected; this is in line with a previous investigation suggesting that human myometrial cells can survive for an extended period of time (several days) under in vitro conditions, regardless of oxygen availability for energy metabolism (15). The release pattern of lactate under strictly anaerobic conditions in the present study revealed a mainly unidirectional flux of this metabolite from the myometrial cells to the extracellular medium during the first 24 h. This first phase of lactate delivery was followed by a second phase, culminating in the attainment of a peak value after 48 h of incubation.…”

Section: Discussionsupporting

confidence: 90%

Lactate distribution in culture medium of human myometrial biopsies incubated under different conditions

Åkerud¹,

Ronquist

Wiberg-Itzel

2009

American Journal of Physiology-Endocrinology and Metabolism

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It is generally believed that a relationship exists between muscle fatigue and intracellular accumulation of lactate. This reasoning is relevant to obstetrical issues. Myocytes in uterus work together during labor, and the contractions need to be strong and synchronized for a child to be delivered. At labor dystocia, the progress of labor becomes slow or arrested after a normal beginning. It has been described that, during labor dystocia, when the force of the contractions is low, the uterus is under hypoxia, and anaerobic conditions with high levels of lactate in amniotic fluid dominate. The purpose of this study was to examine whether myometrial cells are involved in the production of lactate in amniotic fluid and whether there are differences in production and distribution of lactate in cells incubated under aerobic and anaerobic conditions. We also wanted to elucidate the involvement of specific membrane-bound lactate carriers. Women undergoing elective caesarean section were included. Myometrial biopsies from uteri were collected and subjected to either immunohistochemistry to identify lactate carriers or in vitro experiments to analyze production of lactate. The presence of lactate carriers named monocarboxylate transporters 1 and 4 was verified. Myometrial cells produced lactate extracellularly, and the lactate carriers operated differently under anaerobic and aerobic conditions; while being mainly unidirectional under anaerobic conditions, they became bidirectional under aerobic conditions. Human myometrial cells produced and delivered lactate to the extracellular medium under both anaerobic and aerobic conditions. The delivery was mediated by lactate carriers.

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Section: Discussionsupporting

confidence: 90%

Lactate distribution in culture medium of human myometrial biopsies incubated under different conditions

Åkerud¹,

Ronquist

Wiberg-Itzel

2009

American Journal of Physiology-Endocrinology and Metabolism

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What Determines the Intracellular ATP Concentration

2002

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Analysis is made of the mechanisms that control the intracellular ATP level. The balance between energy production and expenditure determines the energy charge of the cell and the ratio of [ATP] to the adenylate pool. The absolute ATP concentration is determined by the adenylate pool, which, in its turn, depends on the balance between the rates of AMP synthesis and degradation. Experimental data are discussed that demonstrate an increase in the adenylate pool in response to activation of energy-consuming processes. A hypothesis is proposed according to which variation in the adenylate pool and absolute ATP concentration affords a cell the possibility of additional control over processes fulfillinguseful work. A mechanism involved in this regulation is described using human erythrocytes as an example. The hypothesis explains why different metabolic pathways (protein and DNA syntheses, polysaccharide synthesis, and lipid synthesis) use different trinucleotides (GTP, UTP, and CTP, respectively) as an energy source. This allows the cell to independently control these metabolic processes by varying the individual nucleotide pools.

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Immortalization and characterization of human myometrial cells from term-pregnant patients using a telomerase expression vector

Soloff

Jeng

Ilies

et al. 2004

Molecular Human Reproduction

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An examination of cellular processes involved in myometrial function has been greatly assisted by the use of human myometrial cells in primary culture. However, these cells can be used only for several passages before they senesce, and responses to various agents change with time in culture. The use of transformed cells is limited, as they can be polynucleated and can lose or gain chromosomes. We have developed three telomerase-immortalized cell lines from term-pregnant human myometrium to eliminate variability between passage numbers and allow genetic manipulations of myometrial cells to fully characterize signal pathways. These cells have a normal karyotype and were verified to be uterine smooth muscle by immunocytochemical staining for smooth muscle cell-specific alpha-actin and high affinity oxytocin antagonist binding sites. The three cell lines and the cells in primary culture from which they were derived were examined by cDNA microarray analysis. Of >10 000 expressed genes, there were consistent changes in the expression of approximately 1% in the three immortalized cell lines. We were unable to detect any significant differences between primary and immortalized cells in signal pathways such as epidermal growth factor-stimulated epidermal growth factor receptor phosphorylation, insulin-stimulated Akt phosphorylation, oxytocin and lysophosphatidic acid-stimulated extracellular signal-regulated kinase 1 and 2 phosphorylation, myosin light chain phosphorylation, and interleukin-1 induction of IkappaBalpha degradation. The immortalized cells should be useful for a range of studies, including high throughput analyses of the effects of environmental agents on the human myometrium.

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Human Uterine Myocytes Retain Their Energy Charge with No Gross Alterations in Morphology for at Least 8 Days when Cultured under Anaerobic Conditions

Cited by 4 publications

References 20 publications

Lactate distribution in culture medium of human myometrial biopsies incubated under different conditions

Lactate distribution in culture medium of human myometrial biopsies incubated under different conditions

What Determines the Intracellular ATP Concentration

Immortalization and characterization of human myometrial cells from term-pregnant patients using a telomerase expression vector

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