Characterization of Hair Follicle Development in Engineered Skin Substitutes

Sriwiriyanont, Penkanok; Lynch, Kaari A.; McFarland, Kevin L.; Supp, Dorothy M.; Boyce, Steven T.

doi:10.1371/journal.pone.0065664

Cited by 44 publications

(29 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The first group (scaffolds seeded with chimeric cells) formed hairs without sebaceous glands, while the third group (scaffolds with human-only cells) formed no follicles or glands. The study showed that sebaceous glands are not required for hair formation when using this protocol (Sriwiriyanont et al 2013).…”

Section: Synthesis Of Hair Follicles and Sebaceous Glandsmentioning

confidence: 87%

Tissue and Organ Regeneration in Adults

Yannas

2015

108

100

View full text Add to dashboard Cite

Section: Synthesis Of Hair Follicles and Sebaceous Glandsmentioning

confidence: 87%

Tissue and Organ Regeneration in Adults

Yannas

2015

108

100

View full text Add to dashboard Cite

“…The creation of hair follicles has recently been successfully completed in skin substitutes, although the methodology behind it is lengthy and tedious[156,157]. Although it may take time for hair follicles stem cells to be incorporated into clinically available skin substitutes, there is hope that at least in vitro trichogenesis is now possible.…”

Section: Components Of a Skin Substitutementioning

confidence: 99%

Methodologies in creating skin substitutes

Nicholas

Jeschke

Amini-Nik

2016

Cell. Mol. Life Sci.

View full text Add to dashboard Cite

The creation of skin substitutes has significantly decreased morbidity and mortality of skin wounds. Although there are still a number of disadvantages of currently available skin substitutes, there has been a significant decline in research advances over the past several years in improving these skin substitutes. Clinically most skin substitutes used are acellular and do not use growth factors to assist wound healing, key areas of potential in this field of research. This article discusses the five necessary attributes of an ideal skin substitute. It comprehensively discusses the three major basic components of currently available skin substitutes: scaffold materials, growth factors, and cells, comparing and contrasting what has been used so far. It then examines a variety of techniques in how to incorporate these basic components together to act as a guide for further research in the field to create cellular skin substitutes with clinically better results.

show abstract

“…The Lef1, a member of the WNT pathway, is a spe cific marker of early regeneration and development processes of the HF (Sriwiriyanont et al, 2013). We did not observe Lef1 expression in coated aggregates (Fig.…”

Section: Dermal Papilla Inhibits Proliferation Of Keratinocytesmentioning

confidence: 74%

Hair follicle regeneration in vitro

Kalabusheva

Vorotelyak

2016

Paleontol. J.

View full text Add to dashboard Cite

MATERIALS AND METHODSPrimary Cell Culture The skin was obtained after face lift surgery from six patients from 20 to 60 years of age, with the informed consent from "The Clinic of Active Lon gevity Institute of Beauty in Arbat." The samples were transported in DMEM medium (PanEko), with addition of gentamicin (RUP "Belmedpreparaty") and glutamine (Gybco). Prior to cell isolation, skin samples were washed with Hank's solution (PanEko), with gentamicin. DP cells were isolated by a series of fermentation and centrifuging, following the technique developed by Wu et al. (2005) and modified by Chermnykh et al. (2010). The cells were cultured in AmnioMAX TM II medium (Gibco). In experiments involving cells pas sages 1-4, passage 12 of DP cells was used for compar isons of their hair inducing abilities.For keratinocytes isolation, the epidermal layer was separated from skin by fermentation in dispase (Gibco) overnight at +4°C. The epidermal sheet was disrupted by trypsinization for 15 minutes. Kerati nocytes suspension was inoculated in DMEM/F12 medium (PanEko) containing 4mM glutamine, 10% fetal bovine serum (FBS) (HyClone), 10 ng/mL EGF (Sigma), 5 mg/mL insulin (Sigma), and 0.25 mg/mL isoproterenol (Sigma). Viral Cell TransfectionLentiviral constructions, bearing red fluorescent protein TagRFP gene, were provided by the Eurogene company. Transfection was performed using cells of the first passage. Cells were transfected in serum free AmnioMAX TM II medium, with addition of poly brene (Sigma) with tenfold excess concentration of viral particles. Mixed CulturesDP cells and keratinocytes were trypsinized by the standard method. To obtain attached culture, DP cells or fibroblasts were mixed with keratinocytes in 1 : 1 proportion, seeded in 48 well plate (Corning) at a concentration of 10 5 cells per well, and cultured for three days.To obtain mixed aggregates, DP cells and kerati nocytes were mixed in 1 : 1 proportion and cultured in a hanging drop at a concentration of 7 × 10 3 cells per aggregate. Cells were cultured for 3 to 14 days.To obtain coated aggregates, DP cell suspension was placed in a hanging drop at a concentration of 3.5 × 10 3 cells per aggregate. After three days in cul ture, aggregates were transferred to preliminary pre pared hanging drops containing 3.5 × 10 3 kerati nocytes and then additionally cultured for 3 to 14 days. ImmunocytochemistryThe attached culture was fixed in 4% paraformal dehyde (PFA) (Riedel de Haen) for 15 min at room temperature (25 o C) and then washed with phosphate saline buffer (PBS) (Gibco). Primary antibodies against Ki67 (Abcam, ab16667) were diluted 1 : 50 in blocking solution consisting of PBS, with addition of 0.1% Triton X 100 (AppliChem), 0.1% Tween 20 (AppliChem), 0.05% bovine serum albumin (Sigma), and sodium azide (Sigma). Several samples were incu bated in blocking solution without antibodies for neg ative secondary antibody control. They were incu Abstract⎯Hair follicles (HF) are skin appendages that develop as a result of the epithelial mesenchymal interactions. T...

show abstract

Characterization of Hair Follicle Development in Engineered Skin Substitutes

Cited by 44 publications

References 35 publications

Tissue and Organ Regeneration in Adults

Tissue and Organ Regeneration in Adults

Methodologies in creating skin substitutes

Hair follicle regeneration in vitro

Contact Info

Product

Resources

About