Isolation and in vitro cultivation of adrenal cells from mice

Friedrich, Linda; Schuster, Maria; Celis, María F. Rubín de; Berger, Ilona; Bornstein, Stefan R.; Steenblock, Charlotte

doi:10.1016/j.xpro.2021.100999

Cited by 4 publications

(3 citation statements)

References 8 publications

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“…There are still not many studies available. The research are related mainly to the adrenals of mice and rats, including in vitro systems where the mostly adrenal development and stress responses were analyzed (Abou Nader et al, 2022; Aşır et al, 2022; Bornstein et al, 2004; Friedrich et al, 2021; Zheng et al, 2022). In wild animals, adrenal gland physiology was studied scarcely by us and others (Barszcz et al, 2016; Ekele et al, 2014; Kotula‐Balak et al, 2023; Olukole et al, 2016; Pawlicki et al, 2023; Sabbi et al, 2020).…”

Section: Discussionmentioning

confidence: 99%

Is membrane androgen and estrogen receptor signaling imperative in the governing function of the adrenal cortex in the Eurasian beaver (Castor fiber L.)?

Lonc,

Hrabia,

Krakowska

et al. 2024

J Exp Zool Pt A

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There is a need to fully know the physiology of Eurasian beaver due to its essential role in environmental homeostasis. However, a “human factor“ impacts this, including stress conditions and environmental pollution. Adrenal glands protect these all. The regulation of endocrine processes by nonclassical androgen and estrogen signaling, the first and fastest control, is still a matter of research. The specific analyses performed here in mature female and male beaver adrenals contained: anatomical and histological examinations, expression and localization of membrane androgen receptor (zinc transporter, Zinc‐ and Iron‐like protein 9; ZIP9) and membrane estrogen receptor coupled with G protein (GPER), and measurement of zinc (Zn2+) and copper (Ca2+) ion levels and corticosterone levels. We revealed normal anatomical localization, size, and tissue histology in female and male beavers, respectively. Equally, ZIP9 and GPER were localized in the membrane of all adrenal cortex cells. The protein expression of these receptors was higher (p < 0.001) in male than female adrenal cortex cells. Similarly, Zn2+ and Ca2+ ion levels were higher (p < 0.05, p < 0.01) in male than female adrenal cortex. The increased corticosterone levels (p < 0.001) were detected in the adrenal cortex of females when compared to males. The present study is the first to report the presence of nonclassical androgen and estrogen signaling and its possible regulatory function in the adrenal cortex of Eurasian beavers. We assume that this first‐activated and fast‐transmitted regulation can be important in the context of the effect of environmental physical and chemical stressors especially on adrenal cortex cells. The beaver adrenals may constitute an additional supplementary model for searching for universal mechanisms of adrenal cortex physiology and diseases.

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Section: Discussionmentioning

confidence: 99%

Is membrane androgen and estrogen receptor signaling imperative in the governing function of the adrenal cortex in the Eurasian beaver (Castor fiber L.)?

Lonc,

Hrabia,

Krakowska

et al. 2024

J Exp Zool Pt A

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“…For RNA preparation, the C57BL/6J-6N strain was used. Urogenital ridges from E9.5 embryos (22 ± 2 somites), adrenal glands from E14.5 embryos and adrenal cortex+capsule 59 from 12-14 weeks old mice were collected and snap frozen until use.…”

Section: Methodsmentioning

confidence: 99%

In vitrodifferentiation of mouse pluripotent stem cells into glucocorticoid-producing adrenocortical cells

Oikonomakos,

Tedesco,

Motamedi

et al. 2023

Preprint

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SummaryDirected differentiation of pluripotent stem cells into specialized cell types represents an invaluable tool for a wide range of applications. Here, we have exploited single-cell transcriptomic data to develop a step-wisein vitrodifferentiation system from mouse embryonic stem cells into adrenocortical cells. We show that during development the adrenal primordium is embedded in an extracellular matrix containing tenascin and fibronectin. Culturing cells on fibronectin during differentiation increased the expression of the steroidogenic marker NR5A1. Furthermore, 3D cultures in the presence of PKA-pathway activators led to the formation of aggregates composed of different cell types expressing adrenal progenitor or steroidogenic markers, including the adrenocortical specific enzymeCyp21a1. Importantly,in vitrodifferentiated cells secreted thezona Fasciculataspecific corticosterone in response to cAMP activators, but not gonadal hormones, thus confirming the specificity of differentiation towards the adrenal lineage.HighlightsMouse adrenal progenitors differentiate in a tenascin/fibronectin-rich extracellular matrixStepwise differentiation of ES cells yields NR5A1+ adrenocortical cells3D aggregates produce glucocorticoids in response to cAMP signalling

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“…Heterozygous Nes-GFP transgenic mice [16] on the C57BL/6N genetic background were generated as described previously [14]. All mouse colonies were maintained under a 12:12 h light/dark cycle and fed ad libitum.…”

Section: Animalsmentioning

confidence: 99%

Development of adrenal 3-dimensional spheroid cultures: potential for the treatment of adrenal insufficiency and neurodegenerative diseases

Steenblock,

Fliedner,

Spinas

et al. 2024

Explor Endocr Metab Dis

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Aim: Regenerative and curative strategies would be desirable for neurodegenerative and adrenal diseases, and multipotent adrenal stem cells are considered as promising biological tools for this purpose. Stem-like cells with the potential to proliferate and differentiate in vivo and in vitro were discovered in both cortex and medulla of the adrenal gland. Previously, it was demonstrated that nestin-positive progenitors in the cortex and medulla, play an important role under stress. In the present study, the cultivation of these cells was optimized and their growth in vitro was characterized. Methods: Primary cells from the adrenal cortex and medulla from Nes-GFP mice were isolated and the in vitro culture conditions promoting the growth of stem and progenitor cells using different 3-dimensional (3D) spheroid culture models were optimized. Results: Both cortical and medullary cells could be cultured for at least one month under several different low-adherence conditions maintaining their viability and potential to differentiate. Medullary cells grew faster than cortical cells. Endothelin did not affect the cultures. Conclusions: Adrenomedullary and adrenocortical nestin-positive progenitor cells can be cultured long-term in 3D cultures maintaining their proliferation and differentiation capabilities. Such multidimensional models can potentially be used for drug screening to develop personalized medicines or for transplantation to treat neurodegenerative disorders or adrenal diseases, such as adrenal insufficiency.

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Isolation and in vitro cultivation of adrenal cells from mice

Cited by 4 publications

References 8 publications

Is membrane androgen and estrogen receptor signaling imperative in the governing function of the adrenal cortex in the Eurasian beaver (Castor fiber L.)?

Is membrane androgen and estrogen receptor signaling imperative in the governing function of the adrenal cortex in the Eurasian beaver (Castor fiber L.)?

In vitrodifferentiation of mouse pluripotent stem cells into glucocorticoid-producing adrenocortical cells

Development of adrenal 3-dimensional spheroid cultures: potential for the treatment of adrenal insufficiency and neurodegenerative diseases

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