Optimizing suction blister epidermal graft technique in the surgical treatment of vitiligo

Dellatorre, Gerson; Bertolini, Wagner; Castro, Caio César Silva de

doi:10.1590/abd1806-4841.20176332

Cited by 9 publications

(11 citation statements)

References 8 publications

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“…Many suction blister studies employ mono-chamber devices, often utilizing syringes with the plungers removed and attached to vacuum pumps [ 13 , 14 ]. These chambers can be cumbersome, cannot assure equal pressure between independent chambers, and/or lack the ability to monitor the pressure during the application.…”

Section: Resultsmentioning

confidence: 99%

Prolonging culture of primary human keratinocytes isolated from suction blisters with the Rho kinase inhibitor Y-27632

et al. 2018

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Keratinocytes are the most abundant cell type in the epidermis. They prevent desiccation and provide immunological and barrier defense against potential pathogens such as Staphylococcus aureus and Candida albicans. The study of this first line of immune defense may be hindered by invasive isolation methods and/or improper culture conditions to support stem cell maintenance and other potential mechanisms contributing to long-term subcultivation in vitro. Primary keratinocytes have been successfully isolated from blister roofs induced by negative pressure, which separates the epidermis from the dermis in vivo in human subjects. This method allows collection of pure epidermal cells without dermal contamination in a minimally invasive manner. However, the isolated keratinocytes differentiate and senesce when cultured in vitro beyond five passages. Here, we present evidence that the Rho kinase (ROCK) inhibitor Y-27632 can be used to effectively increase the proliferative capabilities of keratinocytes isolated using the suction blister method, similar to what has been previously reported for primary keratinocytes isolated using alternative methods. We show that the increase in passage number is directly correlated to delayed differentiation, and that cells passaged long term with the inhibitor retain their ability to stratify in organotypic raft cultures and respond to cytokine treatment; additionally, the late passage cells have a heterogeneous mix of differentiated and non-differentiated cells which may be predicted by a ratio of select differentiation markers. The described method presents a minimally invasive procedure for keratinocyte isolation and prolonged culture that allows analysis of keratinocyte function in both healthy volunteers and patients with dermatologic diseases.

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Section: Resultsmentioning

confidence: 99%

Prolonging culture of primary human keratinocytes isolated from suction blisters with the Rho kinase inhibitor Y-27632

et al. 2018

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“…Many suction blister studies employ mono-chamber devices, often utilizing syringes with the plungers removed and attached to vacuum pumps [12, 13]. These chambers can be cumbersome, cannot assure equal pressure between independent chambers, and/or lack the ability to monitor the pressure during the application.…”

Section: Resultsmentioning

confidence: 99%

Prolonged culture of primary human keratinocytes isolated from suction blisters

Anderson

Sastalla

Earland

et al. 2018

Preprint

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Keratinocytes are the most abundant cell type in the epidermis. They prevent desiccation and provide immunological and barrier defense against potential pathogens such as Staphylococcus aureus and Candida albicans. The study of this first line of immune defense is hindered by invasive isolation methods and insufficient techniques for long-term passage of primary keratinocytes in vitro. Primary keratinocytes have been successfully isolated from blister roofs induced by negative pressure, which separates the epidermis from the dermis in vivo in human subjects. This method allows collection of pure epidermal cells without dermal contamination in a minimally invasive manner. However, the isolated keratinocytes differentiate and senesce when cultured in vitro beyond five passages. Here, we present evidence that the Rho kinase (ROCK) inhibitor Y-27632 can be used to effectively increase the proliferative capabilities of keratinocytes isolated using the suction blister method, similar to what has been previously reported for primary keratinocytes isolated using alternative methods. We show that the increase in passage number is directly correlated to delayed differentiation, and that cells passaged long term with the inhibitor retain their ability to stratify in organotypic raft cultures and respond to cytokine treatment; additionally, the late passage cells have a heterogeneous mix of differentiated and non-differentiated cells which may be predicted by a ratio of select differentiation markers. The described method presents a minimally invasive procedure for keratinocyte isolation and prolonged culture that allows analysis of keratinocyte function in both healthy volunteers and patients with dermatologic diseases.

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“…Blister fluid was utilized for to analyze immune cells in the skin, such as cell recruitment during allergic inflammation because an influx of immune cells into the blister occurred over multiple days after induction of the blister (100). Suction blistering later was used for epidermal grafting, using cells from both the blister roofs and fluid, to treat stable vitiligo lesions (101). More recently, this technique has been used in other contexts to identify cell populations, protein levels of soluble receptors, and secreted growth factors and cytokines (102)(103)(104).…”

Section: Translational Tools For Vitiligo Researchmentioning

confidence: 99%

Translational Research in Vitiligo

Katz

Harris

2021

Front. Immunol.

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Vitiligo is a disease of the skin characterized by the appearance of white spots. Significant progress has been made in understanding vitiligo pathogenesis over the past 30 years, but only through perseverance, collaboration, and open-minded discussion. Early hypotheses considered roles for innervation, microvascular anomalies, oxidative stress, defects in melanocyte adhesion, autoimmunity, somatic mosaicism, and genetics. Because theories about pathogenesis drive experimental design, focus, and even therapeutic approach, it is important to consider their impact on our current understanding about vitiligo. Animal models allow researchers to perform mechanistic studies, and the development of improved patient sample collection methods provides a platform for translational studies in vitiligo that can also be applied to understand other autoimmune diseases that are more difficult to study in human samples. Here we discuss the history of vitiligo translational research, recent advances, and their implications for new treatment approaches.

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Optimizing suction blister epidermal graft technique in the surgical treatment of vitiligo

Cited by 9 publications

References 8 publications

Prolonging culture of primary human keratinocytes isolated from suction blisters with the Rho kinase inhibitor Y-27632

Prolonging culture of primary human keratinocytes isolated from suction blisters with the Rho kinase inhibitor Y-27632

Prolonged culture of primary human keratinocytes isolated from suction blisters

Translational Research in Vitiligo

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