Iterative Three-Dimensional Epidermis Bioengineering and Xenografting to Assess Long-Term Regenerative Potential in Human Keratinocyte Precursor Cells

Fortunel, Nicolas O.; Bouissou-Cadio, Emmanuelle; Coutier, Julien; Martin, Michèle

doi:10.1007/7651_2019_250

Cited by 4 publications

(5 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Samples were submitted to a single exposure to IR at a low dose (50 mGy) or at a higher dose (2 Gy). The next day, irradiated and non-irradiated tissue samples were xenografted onto recipient nude mice ( Figure 1 D) [ 21 ]. This experimental process enabled the study of human epidermis regeneration in an in vivo context up to complete differentiation.…”

Section: Resultsmentioning

confidence: 99%

“…Plasma-based human skin substitutes were reconstructed according to [ 12 , 20 , 21 ]. Human plasma (generous gift from Pr Lataillade, Biomedical Research Institute of French Armies (IRBA), INSERM U1197 Clamart, France), was mixed on ice with 4.68 mg/mL sodium chloride (Fresenius Medical Care, Savigny, France), 0.8 mg/mL CaCl 2 (Laboratoire Renaudin, Itxassou, France), 9.7 μg/mL Exacyl (tranexamic acid) (Sanofi, France) and human dermal fibroblasts.…”

Section: Methodsmentioning

confidence: 99%

“…Experimental procedures [12,21] were approved by the ethical committee CEEA-51 from the Center for Exploration and Experimental Functional Research (CERFE) (Genople ® , Evry, France). Experiments and housing were managed at CERFE under appropriate aseptic conditions.…”

Section: Skin Substitute Xenograftingmentioning

confidence: 99%

See 2 more Smart Citations

Exposure of Human Skin Organoids to Low Genotoxic Stress Can Promote Epithelial-to-Mesenchymal Transition in Regenerating Keratinocyte Precursor Cells

Cavallero

Granito

Stockholm

et al. 2020

Cells

Self Cite

View full text Add to dashboard Cite

For the general population, medical diagnosis is a major cause of exposure to low genotoxic stress, as various imaging techniques deliver low doses of ionizing radiation. Our study investigated the consequences of low genotoxic stress on a keratinocyte precursor fraction that includes stem and progenitor cells, which are at risk for carcinoma development. Human skin organoids were bioengineered according to a clinically-relevant model, exposed to a single 50 mGy dose of γ rays, and then xeno-transplanted in nude mice to follow full epidermis generation in an in vivo context. Twenty days post-xenografting, mature skin grafts were sampled and analyzed by semi-quantitative immuno-histochemical methods. Pre-transplantation exposure to 50 mGy of immature human skin organoids did not compromise engraftment, but half of xenografts generated from irradiated precursors exhibited areas displaying focal dysplasia, originating from the basal layer of the epidermis. Characteristics of epithelial-to-mesenchymal transition (EMT) were documented in these dysplastic areas, including loss of basal cell polarity and cohesiveness, epithelial marker decreases, ectopic expression of the mesenchymal marker α-SMA and expression of the EMT promoter ZEB1. Taken together, these data show that a very low level of radiative stress in regenerating keratinocyte stem and precursor cells can induce a micro-environment that may constitute a favorable context for long-term carcinogenesis.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Exposure of Human Skin Organoids to Low Genotoxic Stress Can Promote Epithelial-to-Mesenchymal Transition in Regenerating Keratinocyte Precursor Cells

Cavallero

Granito

Stockholm

et al. 2020

Cells

Self Cite

View full text Add to dashboard Cite

show abstract

“…Moreover, the regenerative capacity of KLF4 stably repressed keratinocytes was higher than that of KLF4 wild-type keratinocytes. This gain-of-function was demonstrated in vitro by the generation of epidermis organoids, and in vivo by the augmented capacity of KLF4 stably repressed keratinocytes to ensure iterative xenografting in immuno-deficient athymic nude Foxn1 nu mice [ 16 ]. In a first cycle of xenografting, KLF4 repressed and KLF4 wild-type keratinocytes were equally potent for regeneration, indicating an absence of deleterious effects of KLF4 repression on epidermis stratification and differentiation.…”

Section: Klf4 Functions In Native Adult Keratinocytesmentioning

confidence: 99%

When the Search for Stemness Genes Meets the Skin Substitute Bioengineering Field: KLF4 Transcription Factor under the Light

Fortunel

Martin

2020

Cells

Self Cite

View full text Add to dashboard Cite

The transcription factor “Kruppel-like factor 4” (KLF4) is a central player in the field of pluripotent stem cell biology. In particular, it was put under the spotlight as one of the four factors of the cocktail originally described for reprogramming into induced pluripotent stem cells (iPSCs). In contrast, its possible functions in native tissue stem cells remain largely unexplored. We recently published that KLF4 is a regulator of “stemness” in human keratinocytes. We show that reducing the level of expression of this transcription factor by RNA interference or pharmacological repression promotes the ex vivo amplification and regenerative capacity of two types of cells of interest for cutaneous cell therapy: native keratinocyte stem and progenitor cells from adult epidermis, which have been used for more than three decades in skin graft bioengineering, and keratinocytes generated by the lineage-oriented differentiation of embryonic stem cells (ESCs), which have potential for the development of skin bio-bandages. At the mechanistic level, KLF4 repression alters the expression of a large set of genes involved in TGF-β1 and WNT signaling pathways. Major regulators of TGF-β bioavailability and different TGF-β receptors were targeted, notably modulating the ALK1/Smad1/5/9 axis. At a functional level, KLF4 repression produced an antagonist effect on TGFβ1-induced keratinocyte differentiation.

show abstract

“…Keratinocyte holoclones are endowed with the capacity for self-renewal, resulting in a growth potential exceeding 100 population doublings [ 13 , 14 ]. Experimentally, their regenerative potential can be assessed in vitro by their efficient capacity for three-dimensional epidermis reconstruction [ 14 , 15 , 16 ], and in vivo by the xenografting of skin reconstructs in immunocompromised recipient animals [ 17 , 18 , 19 ]. In summary, the cellular model of holoclone keratinocytes has the advantage of connecting basic researches aimed at acquiring knowledge on epidermal stem cells and the clinical side of their uses.…”

Section: Introductionmentioning

confidence: 99%

Deciphering “Immaturity-Stemness” in Human Epidermal Stem Cells at the Levels of Protein-Coding and Non-Coding Genomes: A Prospective Computational Approach

Vinasco-Sandoval,

Lemaître,

Soularue

et al. 2024

IJMS

Self Cite

View full text Add to dashboard Cite

The epidermis hosts populations of epithelial stem cells endowed with well-documented renewal and regenerative functions. This tissue thus constitutes a model for exploring the molecular characteristics of stem cells, which remain to date partially characterized at the molecular level in human skin. Our group has investigated the regulatory functions of the KLF4/TGFB1 and the MAD4/MAX/MYC signaling pathways in the control of the immaturity-stemness versus differentiation fate of keratinocyte stem and precursor cells from human interfollicular epidermis. We described that down-modulation of either KLF4 or MXD4/MAD4 using RNA interference tools promoted an augmented stemness cellular status; an effect which was associated with significant transcriptional changes, as assessed by RNA-sequencing. Here, we have implemented a computational approach aimed at integrating the level of the coding genome, comprising the transcripts encoding conventional proteins, and the non-coding genome, with a focus on long non-coding RNAs (lncRNAs). In addition, datasets of micro-RNAs (miRNAs) with validated functions were interrogated in view of identifying miRNAs that could make the link between protein-coding and non-coding transcripts. Putative regulons comprising both coding and long non-coding transcripts were built, which are expected to contain original pro-stemness candidate effectors available for functional validation approaches. In summary, interpretation of our basic functional data together with in silico biomodeling gave rise to a prospective picture of the complex constellation of transcripts regulating the keratinocyte stemness status.

show abstract

Iterative Three-Dimensional Epidermis Bioengineering and Xenografting to Assess Long-Term Regenerative Potential in Human Keratinocyte Precursor Cells

Cited by 4 publications

References 14 publications

Exposure of Human Skin Organoids to Low Genotoxic Stress Can Promote Epithelial-to-Mesenchymal Transition in Regenerating Keratinocyte Precursor Cells

Exposure of Human Skin Organoids to Low Genotoxic Stress Can Promote Epithelial-to-Mesenchymal Transition in Regenerating Keratinocyte Precursor Cells

When the Search for Stemness Genes Meets the Skin Substitute Bioengineering Field: KLF4 Transcription Factor under the Light

Deciphering “Immaturity-Stemness” in Human Epidermal Stem Cells at the Levels of Protein-Coding and Non-Coding Genomes: A Prospective Computational Approach

Contact Info

Product

Resources

About