Abstract:SummaryCornification and epidermal barrier defects are associated with a number of clinically diverse skin disorders. However, a suitable in vitro model for studying normal barrier function and barrier defects is still lacking. Here, we demonstrate the generation of human epidermal equivalents (HEEs) from human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). HEEs are structurally similar to native epidermis, with a functional permeability barrier. We exposed a pure population of hESC/i… Show more
“…Human neonatal foreskin fibroblasts BJ (ATCC, CRL-2522) were reprogrammed using modified synthetic mRNA as described [10]. At day 2-17 of reprogramming, some cells also were treated with 1 mM pifithrin-a.…”
Section: Reprogrammingmentioning
confidence: 99%
“…At day 2-17 of reprogramming, some cells also were treated with 1 mM pifithrin-a. Two clones with a similar growth rate, one derived in the absence (iKCL004) and one in the presence of 1 mM pifithrin-a (iKCL011), were further characterized; pluripotency marker expression and differentiation into three germ layers in vitro and in vivo (teratomas) revealed no obvious difference between the lines [10].…”
Section: Reprogrammingmentioning
confidence: 99%
“…Differentiation into keratinocytes and generation of HEEs were described previously [10]. Transepithelial electrical resistance was measured with epithelial voltohmmeter EVOM (World Precision Instruments) as described [10,21]. Normal human keratinocytes (NHK) were cultured in EpiLife (Life Technologies).…”
Section: Cell Culturementioning
confidence: 99%
“…A total amount of 750 ng of biotin-labeled aRNA in a 5 mL volume was then used for HumanHT-12 Expression BeadChip whole-genome gene expression direct hybridization assay system (Illumina) according to manufacturer's instructions, run on iScan system (Illumina) and analyzed as described [10]. All samples were analyzed as biological replicates from three independent experiments.…”
“…Two iPSC clones, iKCL004 and iKCL011, that we used to build human skin equivalents [human epidermal equivalent (HEE)] with a functional permeability barrier [10] showed subtle differences, which prompted us to investigate in depth the genetic and epigenetic footprint of both lines. Since the focus of our work was keratinocyte differentiation culminating in the stratum corneum derived epidermal permeability barrier, we concentrated on the epidermal differentiation complex (EDC) on chromosome 1, which contains multiple genes involved in epidermal cornification [11][12][13][14][15][16][17][18][19].…”
“…Human neonatal foreskin fibroblasts BJ (ATCC, CRL-2522) were reprogrammed using modified synthetic mRNA as described [10]. At day 2-17 of reprogramming, some cells also were treated with 1 mM pifithrin-a.…”
Section: Reprogrammingmentioning
confidence: 99%
“…At day 2-17 of reprogramming, some cells also were treated with 1 mM pifithrin-a. Two clones with a similar growth rate, one derived in the absence (iKCL004) and one in the presence of 1 mM pifithrin-a (iKCL011), were further characterized; pluripotency marker expression and differentiation into three germ layers in vitro and in vivo (teratomas) revealed no obvious difference between the lines [10].…”
Section: Reprogrammingmentioning
confidence: 99%
“…Differentiation into keratinocytes and generation of HEEs were described previously [10]. Transepithelial electrical resistance was measured with epithelial voltohmmeter EVOM (World Precision Instruments) as described [10,21]. Normal human keratinocytes (NHK) were cultured in EpiLife (Life Technologies).…”
Section: Cell Culturementioning
confidence: 99%
“…A total amount of 750 ng of biotin-labeled aRNA in a 5 mL volume was then used for HumanHT-12 Expression BeadChip whole-genome gene expression direct hybridization assay system (Illumina) according to manufacturer's instructions, run on iScan system (Illumina) and analyzed as described [10]. All samples were analyzed as biological replicates from three independent experiments.…”
“…Two iPSC clones, iKCL004 and iKCL011, that we used to build human skin equivalents [human epidermal equivalent (HEE)] with a functional permeability barrier [10] showed subtle differences, which prompted us to investigate in depth the genetic and epigenetic footprint of both lines. Since the focus of our work was keratinocyte differentiation culminating in the stratum corneum derived epidermal permeability barrier, we concentrated on the epidermal differentiation complex (EDC) on chromosome 1, which contains multiple genes involved in epidermal cornification [11][12][13][14][15][16][17][18][19].…”
New emerging technologies, remarkably miniaturized 3D organ models and microfluidics, enable simulation of the real in vitro microenvironment ex vivo more closely. There are many fascinating features of innovative organ‐on‐a‐chip (OOC) technology, including the possibility of integrating semipermeable and/or stretchable membranes, creating continuous perfusion of fluids into microchannels and chambers (while maintaining laminar flow regime), embedding microdevices like microsensors, microstimulators, micro heaters, or different cell lines, along with other 3D cell culture technologies. OOC systems are designed to imitate the structure and function of human organs, ranging from breathing lungs to beating hearts. This technology is expected to be able to revolutionize cell biology studies, personalized precision medicine, drug development process, and cancer diagnosis/treatment. OOC systems can significantly reduce the cost associated with tedious drug development processes and the risk of adverse drug reactions in the body, which makes drug screening more effective. The review mainly focus on presenting an overview of the several previously developed OOC systems accompanied by subjects relevant to pharmacy‐, cancer‐, and placenta‐on‐a‐chip. The challenging issues and opportunities related to these systems are discussed, along with a future perspective for this technology.
Although skin grafting is one of the most advanced cell therapy technique, wide application of skin substitutes is hampered by the difficulty in securing sufficient amount of epidermal substitute. Additionally, in understanding the progression of skin aging and disease, and in screening the cosmetic and pharmaceutical products, there is lack of a satisfactory human skin-specific in vitro model. Recently, human embryonic stem cells (hESCs) have been proposed as an unlimited and reliable cell source to obtain almost all cell types present in the human body. This review focuses on the potential off-the-shelf use of hESC-derived keratinocytes for future clinical applications as well as a powerful in vitro skin model to study skin function and integrity, host-pathogen interactions and disease pathogenesis. Furthermore, we discuss the industrial applications of hESC-derived keratinized multi-layer epithelium which provides a human-like test platform for understanding disease pathogenesis, evaluation of new therapeutic modalities and assessment of the safety and efficacy of skin cosmetics and therapeutics. Overall, we conclude that the hESC-derived keratinocytes have great potential for clinical, research and industrial applications.
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