Reconstruction of living bilayer human skin equivalent utilizing human fibrin as a scaffold

Mazlyzam, A.L.; Aminuddin, B S; Fuzina, N H; Norhayati, Moktar; Fauziah, O.; Isa, M R; Saim, Lokman; Ruszymah, B H I

doi:10.1016/j.burns.2006.08.022

Cited by 98 publications

(81 citation statements)

References 19 publications

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“…Our HFSCs showed high viability and were easily enriched from disaggregated dermis, suggesting an alternative technique to cell-sorting systems [17]. Although many types of cells reside in the dermis, for example, sweat gland [18], dermal sheath and dermal papilla [19], nestin [20], endothelial cells [21], melanocytes [22], Merkel cells [23] and fibroblasts [24], serum-free and defined media do not support the growth of unspecific cells and causes these cells to degrade during cultivation. As a result, only HFSCs are sustained in defined media.…”

Section: Isolation and Viability Of Hfscs In Cnt07mentioning

confidence: 99%

A simple culture method for epithelial stem cells derived from human hair follicle

et al. 2013

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The challenge arises among researchers when hair follicle stem cells (HFSCs) derived from a human hair follicle remain poorly expanded in defined culture medium. In this study, we isolated the HFSCs and they became confluent after 10 days of cultivation. Comparing the viability of HFSCs cultured in defined keratinocytes serum free medium (KSFM) in a coated plate and CnT07 medium in an uncoated plate, the number of HFSCs cultured in CnT07 was significantly higher at days 2, 4, 6 and 8 (P=0.004). The population doubling time of HFSCs was 21.48±0.44 hours in non-coated plates with CnT07 and 30.73±0.75 hours in coated plates with KSFM. Our primary HFSC cultures were positive for CD200 and K15 with brownish color. Flow cytometry analysis showed that the percentage of HFSCs expressing CD200 and K15 were 65.20±3.16 and 72.07±6.62 respectively. After reaching 100% confluence, the HFSCs were differentiated into an epidermal layer in vitro using CnT02-3D defined media. HFSCs were differentiated into an epidermal layer after 2 weeks of induction. Involucrin- and K6-positive cells were detected in the differentiated epidermal layer. This method is a simple technique for HFSC isolation and has a lower cost of processing and labor, and it represents a promising tool for skin tissue engineering.

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Section: Isolation and Viability Of Hfscs In Cnt07mentioning

confidence: 99%

A simple culture method for epithelial stem cells derived from human hair follicle

et al. 2013

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“…17 All studies involving human subjects were approved by the ethics committee of Shiraz University of Medical Sciences in accordance with the ethical guidelines of the 1975 Declaration of Helsinki.…”

Section: Methodsmentioning

confidence: 99%

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2015

Exp Clin Transplant

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Objectives: Large-scale expansion of epidermal keratinocytes is essential in the application of these cells for severe burn treatment in patients. Therefore, this study was designed to evaluate various conditions in the expansion of human epidermal keratinocytes. Materials and Methods:The epidermis was separated from the dermis of skin samples using dispase. The epidermis was trypsinized for keratinocyte isolation. Keratinocytes were cultured in various conditions, with or without a human dermal fibroblast feeder layer, mitomycin C treatment, and different culture media. Results: Our results suggest that keratinocytes cultured on a human dermal fibroblast feeder layer were grown for several passages. Extensive deformation and rapid deterioration were observed in the cultured cells without a feeder layer and in serum-free medium. Conclusions: Human dermal fibroblasts treated with mitomycin C can provide optimal conditions for proliferation of keratinocytes.

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“…12 Skin pieces (n = 10) were provided by the plastic surgery department, with informed consent of patients who had reduction mammoplasty. The skin pieces were transferred to the cell culture laboratory in defined keratinocyte serum-free medium (DKSFM) supplemented with 5 μg/mL gentamicin.…”

Section: Keratinocyte Isolationmentioning

confidence: 99%

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2015

Exp Clin Transplant

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Reconstruction of living bilayer human skin equivalent utilizing human fibrin as a scaffold

Cited by 98 publications

References 19 publications

A simple culture method for epithelial stem cells derived from human hair follicle

A simple culture method for epithelial stem cells derived from human hair follicle

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