In vivo transcriptomic profiling using cell encapsulation identifies effector pathways of systemic aging

Mashinchian, Omid; Hong, Xiaotong; Michaud, Joris; Migliavacca, Eugenia; Lefebvre, Grégory; Boss, Christophe; Franceschi, Filippo De; Moal, Eric Le; Collerette-Tremblay, Jasmin; Isern, Joan; Métairon, Sylviane; Raymond, Frédéric; Descombes, Patrick; Bouché, Nicolas; Muñoz‐Cánoves, Pura; Feige, Jérôme N.; Bentzinger, C. Florian

doi:10.7554/elife.57393

Cited by 3 publications

(1 citation statement)

References 56 publications

(68 reference statements)

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“…Previous methods have utilized MuSC transplantation for lineage tracing or regeneration studies (Feige & Rudnicki, 2020) and the encapsulation of MuSCs to determine the effect of systemic factors on stem cells (Mashinchian et al., 2022). MuSC transplantation has been used to verify the capacity of MuSCs to lead to the regeneration of new fibers in injured TA muscles under different experimental conditions (Liu et al., 2018; Price et al., 2014).…”

Section: Introductionmentioning

confidence: 99%

Transcriptional Profiling of Skeletal Muscle Stem Cells After In Vivo Engraftment into a Heterologous Niche Environment

Lazure,

Blackburn,

Soleimani

2023

Current Protocols

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Adult stem cells play a critical role in the maintenance and repair of the organs in which they reside. However, their function is highly dependent on the crosstalk with their niche environment that changes during development and in disease states. The niche provides signals to stem cells to activate, proliferate, self‐renew, or remain in quiescence. In skeletal muscle, the niche is perturbed in disease contexts such as aging, muscular dystrophies, and cachexia. Therefore, it is important to develop methods that permit the decoupling of niche‐mediated from cell‐intrinsic changes that occur in muscle stem cells (MuSCs) in development and disease contexts. With the purpose of determining the effect of the niche environment on the MuSC transcriptome, function, or health, we have coupled an allogeneic stem cell transplantation system, meaning the transplantation of MuSCs from a donor mouse into a recipient host mouse, with Switching Mechanism at 5’ End of RNA Template (SMART‐Seq) to quantify the effects of the niche on the MuSC transcriptome in vivo. Briefly, MuSCs are isolated from a GFP reporter donor mouse (Pax7‐nGFP) and transplanted into the irradiated muscles of immunocompromised allogeneic hosts. The MuSCs are re‐isolated by fluorescence‐activated cell sorting (FACS) after three weeks of inhabiting the heterologous niche, defined as a niche that is different from their originating niche, and sequencing‐ready libraries are created. This method allows for the direct comparison of the transcriptome of stem cells before and after transplantation into a host of a different age, disease status, or genetic background. This method can be used to accurately quantify the direct effect of the niche environment on the stem cell gene expression profile and to decouple cell‐intrinsic versus niche‐mediated alterations in the stem cell transcriptome. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.Basic Protocol: Allogeneic muscle stem cell transplantation

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Section: Introductionmentioning

confidence: 99%

Transcriptional Profiling of Skeletal Muscle Stem Cells After In Vivo Engraftment into a Heterologous Niche Environment

Lazure,

Blackburn,

Soleimani

2023

Current Protocols

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Polymeric Hydrogels for In Vitro 3D Ovarian Cancer Modeling

Braccini

Tacchini

Chiellini

et al. 2022

IJMS

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Ovarian cancer (OC) grows and interacts constantly with a complex microenvironment, in which immune cells, fibroblasts, blood vessels, signal molecules and the extracellular matrix (ECM) coexist. This heterogeneous environment provides structural and biochemical support to the surrounding cells and undergoes constant and dynamic remodeling that actively promotes tumor initiation, progression, and metastasis. Despite the fact that traditional 2D cell culture systems have led to relevant medical advances in cancer research, 3D cell culture models could open new possibilities for the development of an in vitro tumor microenvironment more closely reproducing that observed in vivo. The implementation of materials science and technology into cancer research has enabled significant progress in the study of cancer progression and drug screening, through the development of polymeric scaffold-based 3D models closely recapitulating the physiopathological features of native tumor tissue. This article provides an overview of state-of-the-art in vitro tumor models with a particular focus on 3D OC cell culture in pre-clinical studies. The most representative OC models described in the literature are presented with a focus on hydrogel-based scaffolds, which guarantee soft tissue-like physical properties as well as a suitable 3D microenvironment for cell growth. Hydrogel-forming polymers of either natural or synthetic origin investigated in this context are described by highlighting their source of extraction, physical-chemical properties, and application for 3D ovarian cancer cell culture.

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In vivo transcriptomic profiling using cell encapsulation identifies effector pathways of systemic aging

Mashinchian

Hong

Michaud

et al. 2022

eLife

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Sustained exposure to a young systemic environment rejuvenates aged organisms and promotes cellular function. However, due to the intrinsic complexity of tissues it remains challenging to pinpoint niche-independent effects of circulating factors on specific cell populations. Here we describe a method for the encapsulation of human and mouse skeletal muscle progenitors in diffusible polyethersulfone hollow fiber capsules that can be used to profile systemic aging in vivo independent of heterogeneous short-range tissue interactions. We observed that circulating long-range signaling factors in the old systemic environment lead to an activation of Myc and E2F transcription factors, induce senescence and suppress myogenic differentiation. Importantly, in vitro profiling using young and old serum in 2D culture does not capture all pathways deregulated in encapsulated cells in aged mice. Thus, in vivo transcriptomic profiling using cell encapsulation allows for the characterization of effector pathways of systemic aging with unparalleled accuracy.

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In vivo transcriptomic profiling using cell encapsulation identifies effector pathways of systemic aging

Cited by 3 publications

References 56 publications

Transcriptional Profiling of Skeletal Muscle Stem Cells After In Vivo Engraftment into a Heterologous Niche Environment

Transcriptional Profiling of Skeletal Muscle Stem Cells After In Vivo Engraftment into a Heterologous Niche Environment

Polymeric Hydrogels for In Vitro 3D Ovarian Cancer Modeling

In vivo transcriptomic profiling using cell encapsulation identifies effector pathways of systemic aging

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