Isolation of Stem Cells and Progenitors from Mouse Epidermis

Kostic, Lana; Sedov, Egor; Soteriou, Despina; Yosefzon, Yahav; Fuchs, Yaron

doi:10.1002/cpsc.26

Cited by 9 publications

(5 citation statements)

References 35 publications

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“…Underlying adipose tissue was removed before incubting in trypsin overnight at 4 °C or 2 h at 37 °C 42 . Epidermis and hairs were collected and filtered before sorting 43 . Isolation of α6 + ScaI − Blimp-YFP + cells was performed using a BD FACSAria IIIu sorter, sorting for endogenous Blimp1 -YFP + and utilizing antibodies against Integrin α6 (eBioscience, #12-0495-82) and ScaI (BD Pharmigen, #558162).…”

Section: Methodsmentioning

confidence: 99%

Blimp1+ cells generate functional mouse sebaceous gland organoids in vitro

et al. 2019

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Most studies on the skin focus primarily on the hair follicle and interfollicular epidermis, whereas little is known regarding the homeostasis of the sebaceous gland (SG). The SG has been proposed to be replenished by different pools of hair follicle stem cells and cells that resides in the SG base, marked by Blimp1. Here, we demonstrate that single Blimp1 + cells isolated from mice have the potential to generate SG organoids in vitro. Mimicking SG homeostasis, the outer layer of these organoids is composed of proliferating cells that migrate inward, undergo terminal differentiation and generating lipid-filled sebocytes. Performing confocal microscopy and mass-spectrometry, we report that these organoids exhibit known markers and a lipidomic profile similar to SGs in vivo. Furthermore, we identify a role for c-Myc in sebocyte proliferation and differentiation, and determine that SG organoids can serve as a platform for studying initial stages of acne vulgaris, making this a useful platform to identify potential therapeutic targets.

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Section: Methodsmentioning

confidence: 99%

Blimp1+ cells generate functional mouse sebaceous gland organoids in vitro

et al. 2019

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“…For hematopoietic stem cells, bone marrow cells were stained using BD Stemflow™ Mouse Hematopoietic Stem and Progenitor Cell Isolation Kit (BD, 560492) following the manufacturer’s instructions. For skin stem cells, populations of interest based on negative DAPI (Fisher Scientific, 5748) expression and surface marker expressions ITGA6 (Invitrogen, 12-0495-82), ITGB1(BioLegend, 102215), CD34 (Invitrogen, 11-0341-82), Sca1 (BioLegend, 122513) were used (74). Flow cytometry data was analyzed using FlowJo software.…”

Section: Methodsmentioning

confidence: 99%

Inactivation of a non-canonical gp130 signaling arm attenuates chronic systemic inflammation and multimorbidity induced by a high-fat diet

Lee,

Sarkar,

Tassey

et al. 2024

Preprint

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Interleukin-6 (IL-6) is a major pro-inflammatory cytokine for which the levels in plasma demonstrate a robust correlation with age and body mass index (BMI) as part of the senescence-associated secretory phenotype. IL-6 cytokines also play a crucial role in metabolic homeostasis and regenerative processes, primarily via the canonical STAT3 pathway. Thus, selective modulation of IL-6 signaling may offer a unique opportunity for therapeutic interventions. Recently, we discovered that a non-canonical signaling pathway downstream of tyrosine (Y) 814 within the intracellular domain of gp130, the IL-6 co-receptor, is responsible for the recruitment and activation of SRC family of kinases (SFK). Mice with constitutive genetic inactivation of gp130 Y814 (F814 mice) show accelerated resolution of inflammatory response and superior regenerative outcomes in skin wound healing and posttraumatic models of osteoarthritis. The current study was designed to explore if selective genetic or pharmacological inhibition of the non-canonical gp130-Y814/SFK signaling reduces systemic chronic inflammation and multimorbidity in a high-fat diet (HFD)-induced model of accelerated aging. F814 mice showed significantly reduced inflammatory response to HFD in adipose and liver tissue, with significantly reduced levels of systemic inflammation compared to wild type mice. F814 mice were also protected from HFD-induced bone loss and cartilage degeneration. Pharmacological inhibition of gp130-Y814/SFK in mice on HFD mirrored the effects observed in F814 mice on HFD; furthermore, this pharmacological treatment also demonstrated a marked increase in physical activity levels and protective effects against inflammation-associated suppression of neurogenesis in the brain tissue compared to the control group. These findings suggest that selective inhibition of SFK signaling downstream of gp130 receptor represents a promising strategy to alleviate systemic chronic inflammation. Increased degenerative changes and tissue senescence are inevitable in obese and aged organisms, but we demonstrated that the systemic response and inflammation-associated multi-morbidity can be therapeutically mitigated.

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“…Hoechst 33342 was added for live cell discrimination. Skin cells were isolated using a modified version of the protocol by Kostic and colleagues ( Kostic et al., 2017 ). Briefly, the dorsal skin of mice was dissected and the connective tissues and fat were removed.…”

Section: Methodsmentioning

confidence: 99%

Visualization of individual cell division history in complex tissues using iCOUNT

et al. 2021

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Summary The division potential of individual stem cells and the molecular consequences of successive rounds of proliferation remain largely unknown. Here, we developed an inducible cell division counter (iCOUNT) that reports cell division events in human and mouse tissues in vitro and in vivo . Analyzing cell division histories of neural stem/progenitor cells (NSPCs) in the developing and adult brain, we show that iCOUNT can provide novel insights into stem cell behavior. Further, we use single-cell RNA sequencing (scRNA-seq) of iCOUNT-labeled NSPCs and their progenies from the developing mouse cortex and forebrain-regionalized human organoids to identify functionally relevant molecular pathways that are commonly regulated between mouse and human cells, depending on individual cell division histories. Thus, we developed a tool to characterize the molecular consequences of repeated cell divisions of stem cells that allows an analysis of the cellular principles underlying tissue formation, homeostasis, and repair.

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Isolation of Stem Cells and Progenitors from Mouse Epidermis

Cited by 9 publications

References 35 publications

Blimp1+ cells generate functional mouse sebaceous gland organoids in vitro

Blimp1+ cells generate functional mouse sebaceous gland organoids in vitro

Inactivation of a non-canonical gp130 signaling arm attenuates chronic systemic inflammation and multimorbidity induced by a high-fat diet

Visualization of individual cell division history in complex tissues using iCOUNT

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