Cell motion predicts human epidermal stemness

Nanba, Daisuke; Toki, Fujio; Tate, Sota; Imai, Masayuki; Matsushita, Natsuki; Shiraishi, Ken; Sayama, Koji; Toki, H.; Higashiyama, Shigeki; Barrandon, Yann

doi:10.1083/jcb.201409024

Cited by 40 publications

(58 citation statements)

References 64 publications

(113 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…18,36 It is also of interest to investigate the extent to which cell migration contributes to patterning on our substrates, since it has recently been reported that epidermal stem cells undergo collective cell movements that are correlated with their self-renewal ability. 37…”

Section: Discussionmentioning

confidence: 99%

Mimicking the topography of the epidermal–dermal interface with elastomer substrates

et al. 2016

View full text Add to dashboard Cite

In human skin the interface between the epidermis and dermis is not flat, but undulates. The dimensions of the undulations change as a function of age and disease. Epidermal stem cell clusters lie in specific locations relative to the undulations; however, whether their location affects their properties is unknown. To explore this, we developed a two-step protocol to create patterned substrates that mimic the topographical features of the human epidermal-dermal interface. Substrates with negative patterns were first fabricated by exposing a photocurable formulation to light, controlling the topographical features (such as diameter, height and center-to-center distance) by the photomask pattern dimensions and UV crosslinking time. The negative pattern was then translated to PDMS elastomer to fabricate substrates with 8 unique surface topographies on which primary human keratinocytes were cultured. We found that cells were patterned according to topography, and that separate cues determined the locations of stem cells, differentiated cells and proliferating cells. The biomimetic platform we have developed will be useful for probing the effect of topography on stem cell behaviour.

show abstract

Section: Discussionmentioning

confidence: 99%

Mimicking the topography of the epidermal–dermal interface with elastomer substrates

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Keratinocytes on 100-mm-diameter islands undergo concerted circular motion; however, inhibition of coordinated cell movement does not interfere with epidermis assembly (Gautrot et al, 2012). This contrasts with the observation that human epidermal stem cells in culture undergo collective cell movements that are correlated with their self-renewal ability, the cells with the highest capacity for self-renewal displaying a unique rotational movement that can be identified as early as the twocell stage colony (Nanba et al, 2015). While the major advantages of the microepidermis platform are uniformity and speed of assembly, currently it does not lend itself to analysis of different stem cell subpopulations that differ in expression of specific markers (Tan et al, 2013a) or properties such as cell movement or balanced versus expanding cell growth (Roshan et al, 2016).…”

Section: Integration Of Different Signals At Cell-cell Bordersmentioning

confidence: 82%

Engineered Microenvironments to Direct Epidermal Stem Cell Behavior at Single-Cell Resolution

Watt

2016

Developmental Cell

View full text Add to dashboard Cite

Mammalian epidermis is maintained through proliferation of stem cells and differentiation of their progeny. The balance between self-renewal and differentiation is controlled by a variety of interacting intrinsic and extrinsic factors. Although the nature of these interactions is complex, they can be modeled in a reductionist fashion by capturing single epidermal stem cells on micropatterned substrates and exposing them to individual stimuli, alone or in combination, over defined time points. These studies have shown that different extrinsic stimuli trigger a common outcome-initiation of terminal differentiation-by activating different signaling pathways and eliciting different transcriptional responses.

show abstract

“…Spheroid-forming cells are likely present in all HKc/HPV16 lines, including NF/ HPV16 Δneg , but are continuously selected for during clonal passaging, thereby enabling them to achieve greater homogeneity and yielding a spheroid-forming phenotype as cells undergo in vitro progression. Given that SD-NHKc are actively proliferating populations of keratinocytes, their rapid clonal divisions may uniquely predispose them to HPV-mediated immortalization, as suggested by recent findings (40,41). Moreover, the presence of hyperchromatic cells within HKc/DR aggregates indicates that they are further along the preneoplastic progression process than their HKc/HPV16 counterparts.…”

Section: Discussionmentioning

confidence: 95%

Stem Cell Properties of Normal Human Keratinocytes Determine Transformation Responses to Human Papillomavirus 16 DNA

Woappi

Hosseinipour

Creek

et al. 2018

J Virol

View full text Add to dashboard Cite

Human papillomavirus (HPV) infection of the genital tract is common; however, only about 10 to 15% of infections persist, and approximately 10 to 15% of these persistent infections result in cancer. Basal epidermal stem cells are the presumed target cells for HPV infection, providing a reservoir of latently infected cells that persist over time and initiate lesions. However, it is not known whether stem cell density has any influence on transformation of human keratinocytes by HPV. We explored the relationship between stem cell properties of normal human keratinocytes and their susceptibility to transformation by HPV16 DNA. Normal human keratinocyte isolates (NHKc) derived from different donors were cultured in three-dimensional anchorage-free suspension to assess their spheroid-forming ability. NHKc spheroids were then plated back into plastic monolayer culture and transfected with full-length HPV16 DNA, which we have previously shown to integrate into the host cell genome upon transfection. Spheroid-derived NHKc (SD-NHKc) and fluorescence-activated cell sorting-purified populations of basal stem-like keratinocytes, expressing low levels of epidermal growth factor receptor and high levels of integrin alpha 6 (EGFR/ITGα6), responded to transfection with HPV16 DNA with more vigorous proliferation, greater immortalization efficiency, and faster progression to differentiation resistance than autologous mass-cultured cells. Conversely, cells committed to terminal differentiation (EGFR/ITGα6) grew slowly after transfection with HPV16 and failed to generate immortalized or DR clones. HPV16 DNA induced stem cell properties in mass-cultured NHKc. We conclude that HPV16 preferentially immortalizes basal keratinocytes with stem cell properties and that these cells readily achieve a differentiation-resistant phenotype upon immortalization by HPV16. This paper explores the relationship between the stem cell properties of normal human epidermal cells in culture and these cells' susceptibility to transformation by HPV16 DNA, the HPV type present in about 50% of cervical cancers. We report variable susceptibilities to HPV16-mediated transformation among different keratinocyte isolates derived from neonatal foreskin. Our findings provide strong experimental evidence that HPV16 preferentially transforms basal keratinocytes with stem cell properties. Insights gained from these studies increase our understanding of the host cell-specific factors influencing individual susceptibility to HPV-driven transformation and the contributing factors leading to preneoplastic and neoplastic progression of HPV-positive lesions.

show abstract

Cell motion predicts human epidermal stemness

Abstract: Keratinocyte stem cell colonies can be identified by analyzing cell motion, an emergent stem cell property.

Cited by 40 publications

References 64 publications

Mimicking the topography of the epidermal–dermal interface with elastomer substrates

Mimicking the topography of the epidermal–dermal interface with elastomer substrates

Engineered Microenvironments to Direct Epidermal Stem Cell Behavior at Single-Cell Resolution

Stem Cell Properties of Normal Human Keratinocytes Determine Transformation Responses to Human Papillomavirus 16 DNA

Contact Info

Product

Resources

About