Serum starvation affects mitochondrial metabolism of adipose-derived stem/stromal cells

Giannasi, Chiara; Niada, Stefania; Morte, Elena Della; Casati, Silvia Rosanna; Palma, Clara De; Brini, A.T.

doi:10.1016/j.jcyt.2023.03.004

Cited by 4 publications

(1 citation statement)

References 44 publications

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“…Starvation alone is usually performed after expansion in FBS or FBS with priming stimuli when the whole secretome or EVs are collected at the price of possible inflammation reduction caused by cytokine removal, while starvation supplemented with inflammation may be envisioned when only EVs at their maximum empowerment have to be produced. A major issue that is often underestimated is the effect of starvation on the cells [37][38][39], often leading researchers to attribute the same features to FBS-fed and starved cells. For this reason, a direct comparison of cells cultured in FBS or starvation is mandatory, and the selection of the most appropriate HKG is crucial.…”

Section: Discussionmentioning

confidence: 99%

Inflammation and Starvation Affect Housekeeping Gene Stability in Adipose Mesenchymal Stromal Cells

Ragni,

Piccolo,

Taiana

et al. 2024

CIMB

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Due to the scientific success of in vitro and in vivo model studies, the interest in using mesenchymal stromal cells (MSCs) for the treatment of orthopaedic conditions is growing. In the context of osteoarthritis (OA), MSCs, and, in particular, those derived from adipose tissues (ASCs), have found broader access to clinical use as active components of minimally manipulated orthobiologics, as well as clinically expanded cell preparations, or to collect their released factors (secretome) for cell-free approaches. In this regard, while both inflammatory priming and starvation are common strategies used to empower cell potency or collect the secretome, respectively, little is known about the possible influence of these approaches on the stability of housekeeping genes (HKGs) for molecular studies able to fingerprint cell phenotype or potency. In this report, the reliability of five commonly used HKGs (ACTB, B2M, GAPDH, HPRT1 and RPLP0) was tested in ASCs cultured under standard protocol after inflammatory priming or starvation. Gene expression data were computed with four different applets able to rank genes depending on their stability in either single or combined conditions. The obtained final ranking suggests that for each treatment, a specific HKG is needed, and that starvation is the condition with the stronger effect on HKGs’ stability and, therefore, reliability. The normalization effect of proper HKGs’ use was then validated on three genes involved in OA and whose product is released by ASCs. Overall, data presented herein confirm that the choice of the best HKG has to be carefully considered and that each specific condition has to be tested to identify the most reliable candidate.

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Section: Discussionmentioning

confidence: 99%

Inflammation and Starvation Affect Housekeeping Gene Stability in Adipose Mesenchymal Stromal Cells

Ragni,

Piccolo,

Taiana

et al. 2024

CIMB

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Benzylidenemalononitrile bridged long-wavelength emission luminogens with “washing-free” properties for monitoring the dynamics of LDs

Shi,

Zhao,

et al. 2025

Dyes and Pigments

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Methods of karyoplast cell cycle synchronization for increasing the efficiency of somatic cloning of farm animals

Zhukova

2024

Genes & Cells

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This article is aimed to the analysis of methods for synchronizing the cell cycle of the somatic cells (karyoplasts) upon somatic cloning of farm animals. To obtain cloned animals, the oocytes at the metaphase of the second meiotic division without prior activation are applied as recipient cells in most experiments. In most experiments to obtain cloned offspring, oocytes are used as recipient cells at metaphase of the second meiotic division without prior activation, which determines the choice of somatic cells as karyoplasts at the G0/G1 stage, the most optimal for subsequent nuclear reprogramming by oocyte cytoplasmic factors. The most commonly used methods for arresting cells in this phase are serum starvation and/or contact inhibition, which allows up to 90% of cells to be stopped at the G0/G1 stage. However, despite the effectiveness of these methods, they have a number of significant limitations, and therefore the addition of components to the culture medium of somatic cells that prevent the progression of cells through the cell cycle stages is becoming widespread. The advantage of using chemical inhibitors is the ability of a number of them to have a protective effect on somatic cells, preventing the induction of apoptotic changes. The efficacy of butyrolactone-I, mimosine and aphidicolin application is controversial but a number of studies attests the possible using of these drugs to synchronize the karyoplasts. Thus, there is a wide range of methods for effective synchronization of the cell cycle of karyoplasts, and when choosing the optimal method, one should pay attention to the type of cells, the species of animals from which they were obtained, the permissible duration of cultivation under given conditions in order to minimize the negative impact of conditions on karyoplast viability.

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Serum starvation affects mitochondrial metabolism of adipose-derived stem/stromal cells

Cited by 4 publications

References 44 publications

Inflammation and Starvation Affect Housekeeping Gene Stability in Adipose Mesenchymal Stromal Cells

Inflammation and Starvation Affect Housekeeping Gene Stability in Adipose Mesenchymal Stromal Cells

Benzylidenemalononitrile bridged long-wavelength emission luminogens with “washing-free” properties for monitoring the dynamics of LDs

Methods of karyoplast cell cycle synchronization for increasing the efficiency of somatic cloning of farm animals

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