Cryopreserved placental biopsies maintain mitochondrial activity for high-resolution respirometry

Giovarelli, Matteo; Serati, Anaïs; Zecchini, Silvia; Guelfi, Fabiola; Clementi, Emilio; Mandò, Chiara

doi:10.1186/s10020-023-00645-2

Cited by 2 publications

References 28 publications

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Impact of cryopreservation on immune cell metabolism as measured by SCENITH

Luscombe,

Jones,

Gregorova

et al. 2024

Preprint

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The dynamic functioning of immune cells is regulated by cellular metabolic processes, and there is growing interest in the study of immunometabolic correlates of dysfunctional immune responses. SCENITH is a novel flow cytometry-based technique that allows forex vivometabolic profiling of immune cells within heterogeneous samples. Cryopreservation of clinical samples is frequently undertaken to facilitate high throughput processing, but is thought to lead to cellular metabolic dysfunction. We aimed to investigate the impact of cryopreservation on immune cell metabolism, harnessing SCENITH’s unique ability to describe the divergent bioenergetic characteristics of distinct immune cell subsets.We demonstrate that T cells undergoing activation with a CD3/CD28 stimulus are less readily metabolically reprogrammed following cryopreservation. Additionally, we find that cryopreservation introduces a time-dependent metabolic artefact that favours glycolysis and impairs oxidative phosphorylation, suggesting that cryopreservation results in mitochondrial dysfunction. Despite this artefact, SCENITH was still able to reveal the distinct bioenergetic profiles of contrasting immune cells populations following cryopreservation – for example, non-classical monocytes have a higher mitochondrial dependence than classical monocytes, and T cell CD69 expression is associated with an upregulation of glycolytic capacity.Whilst we believe that SCENITH can provide valuable information about immune cell metabolism even in cryopreserved samples, our findings have important implications for the design of future studies. Investigators should carefully consider how to process and store clinical samples to ensure that cryopreservation does not confound analyses, particularly where longitudinal sampling is required.

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Impact of cryopreservation on immune cell metabolism as measured by SCENITH

Luscombe,

Jones,

Gregorova

et al. 2024

Preprint

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Placental Bioenergetics and Antioxidant Homeostasis in Maternal Obesity and Gestational Diabetes

Mandò,

Castiglioni,

Novielli

et al. 2024

Antioxidants

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Maternal obesity has been associated with short- and long-term risks of pregnancy-perinatal adverse events, possibly due to alterations of placental mitochondrial bioenergetics. However, several detrimental mechanisms occurring in the placentas of women with obesity still need to be clarified. Here, we analyzed placental mitochondrial features and oxidative environment of 46 pregnancies in relation to pre-pregnancy BMI. Seventeen Caucasian normal-weight (NW) and twenty-nine women who were obese (OB) were enrolled. The protein expression of mitochondrial CypD and electron transfer chain complexes (C) I–V were measured, as well as ATP production and oxygen consumption rates (OCRs). The protein levels of the pro/anti-oxidant enzymes TXNIP, SOD2, and PON2 were also analyzed. Despite no differences in CypD expression, OCRs were significantly lower in OB vs. NW women. Accordingly, ATP synthase (CV) levels and ATP content were decreased in OB women, positively correlating with placental efficiency, suggesting a link between ATP deficiency and placental dysfunction. SOD2 expression negatively correlated with maternal BMI, indicating a possible impairment of antioxidant defenses with increasing BMI. These changes were worsened in 10 OB women presenting with gestational diabetes mellitus. Overall, these results suggest alterations of placental bioenergetics in pregnancies of women with obesity, possibly leading to placental dysfunction and altered fetal development and programming.

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Cryopreserved placental biopsies maintain mitochondrial activity for high-resolution respirometry

Cited by 2 publications

References 28 publications

Impact of cryopreservation on immune cell metabolism as measured by SCENITH

Impact of cryopreservation on immune cell metabolism as measured by SCENITH

Placental Bioenergetics and Antioxidant Homeostasis in Maternal Obesity and Gestational Diabetes

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