Immortalization of Primary Keratinocytes and Its Application to Skin Research

Choi, Moonju; Lee, Choong-Ho

doi:10.4062/biomolther.2015.038

Cited by 37 publications

(37 citation statements)

References 74 publications

(103 reference statements)

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“…While culture of murine keratinocytes has been widely reported, these approaches are typically hampered by at least one of the following features: spontaneous immortalization, dependence on feeder cells, and serum or incomplete differentiation, or limited establishment from adult tissue (20)(21)(22)(23)(24). With the current culture system, we show the presence of proliferative basal layer cells and differentiation toward the cornified envelope.…”

Section: Discussionmentioning

confidence: 91%

“…It has previously been reported that cultured murine keratinocytes senesce over time, immortalize through oncogenic mutations, or acquire increased chromosome numbers (20)(21)(22). We therefore performed karyotyping on late-passage organoids (∼P30).…”

Section: Whole-transcriptome Analysis Identifies Epidermal Organoids mentioning

confidence: 99%

“…A large amount of knowledge on the biology of mammalian epidermal stem cells has been generated using rodent models (1,4,5,10). However, in culture, murine epidermal keratinocytes spontaneously immortalize, have limited differentiation capacity, and require feeder cells or serumcontaining conditions, or can be only solidly established from neonatal skin (20)(21)(22)(23)(24). Thus, a long-term in vitro system for the expansion and differentiation of murine epidermal stem cells currently does not exist.…”

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confidence: 99%

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Long-term expansion and differentiation of adult murine epidermal stem cells in 3D organoid cultures

Boonekamp

Kretzschmar

Wiener

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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Mammalian epidermal stem cells maintain homeostasis of the skin epidermis and contribute to its regeneration throughout adult life. While 2D mouse epidermal stem cell cultures have been established decades ago, a long-term, feeder cell- and serum-free culture system recapitulating murine epidermal architecture has not been available. Here we describe an epidermal organoid culture system that allows long-term, genetically stable expansion of adult epidermal stem cells. Our epidermal expansion media combines atypically high calcium concentrations, activation of cAMP, FGF, and R-spondin signaling with inhibition of bone morphogenetic protein (BMP) signaling. Organoids are established robustly from adult mouse skin and expand over at least 6 mo, while maintaining the basal-apical organization of the mouse interfollicular epidermis. The system represents a powerful tool to study epidermal homeostasis and disease in vitro.

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Section: Discussionmentioning

confidence: 91%

Section: Whole-transcriptome Analysis Identifies Epidermal Organoids mentioning

confidence: 99%

mentioning

confidence: 99%

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Long-term expansion and differentiation of adult murine epidermal stem cells in 3D organoid cultures

Boonekamp

Kretzschmar

Wiener

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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“…E7 protein is known to bind with multiple targets specifically RB to overcome the restriction points [41]. Primary keratinocytes get immortalized due to ectopic expression of E6 and E7 which also induces the genomic instability [42]. Carcinogenesis is the consequence of viral gene expression, dysregulated cell proliferation, and genomic instability; however, the progression of cancer is not beneficial to the virus.…”

Section: Hpv Genome and Replicationmentioning

confidence: 99%

Recent Advances in Human Papillomavirus Infection and Management

Saxena¹,

Kumar²,

Goel³

et al. 2019

Current Perspectives in Human Papillomavirus

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Human papillomavirus (HPV) accounts for approximately 4.5% of all cancers which differs at the level of economic development and geographical regions. The life cycle of the HPV is completely dependent on the epithelium differentiation without the involvement of cell death and systemic viremia. Carcinogenesis is the consequence of viral gene expression, dysregulated cell proliferation, and genomic instability. Keratinocytes are the target cell for HPV which act as the physical and immunological barrier. In cervical carcinogenesis, the enhanced level of Th17 infiltration has been observed which increases with the disease progression and is coupled with CCL20 expression in the stromal mesenchymal compartment. IL-6 and M-CSF are known as "switch factors" which are imperative for pro-tumorigenic response in monocytes. Screening of cervical cancer includes three major procedures: cytology, nucleic acid test, and co-testing. For evaluating anal lesions, high-resolution anoscopy is performed which is similar to colposcopy. Prophylactic vaccination is the primary preventive measure to control the HrHPV infection and reduce the burden of HPV-related cancer. The precancerous stage of HPV infection includes excision, ablation, and immunotherapy. Radiotherapy is the acceptable primary treatment for the early stage of anogenital cancer, whereas for the advanced-stage metastatic cancer, palliative therapy is the only option.

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“…In the present study, the A431 (NMSC) cell line and a nontumorigenic immortalized keratinocyte (HaCaT) cell line were examined. HaCaT cells predominantly exist in the basal layer of the skin (Choi & Lee, ) and were used as the control treatment. The aim of the present study was to investigate the effects of cristazine (Figure a) on apoptosis and cell growth inhibition in human epidermoid carcinoma (A431) cells.…”

Section: Introductionmentioning

confidence: 99%

Cristazine, a novel dioxopiperazine alkaloid, induces apoptosis via the death receptor pathway in A431 cells

Patil

Jung

et al. 2019

Drug Development Research

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The fungus Chaetomium sp. is a causative agent of infections in humans and is ubiquitous in the natural environment. The secondary metabolites of this genus exhibit many biological activities, including antifungal activity and toxicity in mitochondria. In this study, we isolated cristazine from the fungus C. cristatum, which has the potential to inhibit the growth of human epidermoid carcinoma (A431) cells in a dose-and time-dependent manner. Its inhibitory activity was examined using a cell viability assay and cell death was elucidated by western blot analysis. Cristazine triggered apoptotic cell death via the Type I death receptor pathway including the activation of caspases and other target proteins. However, cristazine did not have any effect on mitochondrial apoptotic proteins such as Bid, cytochrome c, and apoptosis-inducing factor. Cristazine inhibited the cell cycle progression by arresting the G 1 /S phase and up-regulating the inhibitory proteins of cyclin-dependent kinases. Thus, cristazine has great potential for inducing apoptosis in A431 cells via both cell cycle arrest and the inhibition of cell growth. K E Y W O R D S cell cycle arrest, Chaetomium cristatum, cristazine, death receptor apoptosis, human epidermoid carcinoma

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Immortalization of Primary Keratinocytes and Its Application to Skin Research

Cited by 37 publications

References 74 publications

Long-term expansion and differentiation of adult murine epidermal stem cells in 3D organoid cultures

Long-term expansion and differentiation of adult murine epidermal stem cells in 3D organoid cultures

Recent Advances in Human Papillomavirus Infection and Management

Cristazine, a novel dioxopiperazine alkaloid, induces apoptosis via the death receptor pathway in A431 cells

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