Mitomycin C-Treated 3T3 Fibroblasts Used as Feeder Layers for Human Keratinocyte Culture Retain the Capacity to Generate Eicosanoids

Blacker, Kerry L.; Williams, Mary L.; Goldyne, Marc E.

doi:10.1111/1523-1747.ep12461169

Cited by 29 publications

(12 citation statements)

References 7 publications

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“…Indeed, fibroblast-generated PGE 2 is known to influence keratinocyte proliferation (25). Furthermore, when culturing keratinocytes, 3T3 fibroblasts treated with mitomycin c are sometimes used as a feeder line to provide the required growth factors including eicosanoids, indicating the relevance for the keratinocyte-fibroblast interaction (26). Thus, any disruptions in the eicosanoid generation potential of the fibroblasts will most likely affect the keratinocyte proliferation, compromising the epithelialization step of the wound-healing process.…”

Section: Discussionmentioning

confidence: 99%

Ceramide kinase is required for a normal eicosanoid response and the subsequent orderly migration of fibroblasts

Wijesinghe

Brentnall

Mietla

et al. 2014

Journal of Lipid Research

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In these studies, the role of ceramide-1-phosphate (C1P) in the wound-healing process was investigated. Specifically, fibroblasts isolated from mice with the known anabolic enzyme for C1P, ceramide kinase (CERK), ablated (CERK−/− mice) and their wild-type littermates (CERK+/+) were subjected to in vitro wound-healing assays. Simulation of mechanical trauma of a wound by scratching a monolayer of fibroblasts from CERK+/+ mice demonstrated steadily increasing levels of arachidonic acid in a time-dependent manner in stark contrast to CERK−/− fibroblasts. This observed difference was reflected in scratch-induced eicosanoid levels. Similar, but somewhat less intense, changes were observed in a more complex system utilizing skin biopsies obtained from CERK-null mice. Importantly, C1P levels increased during the early stages of human wound healing correlating with the transition from the inflammatory stage to the peak of the fibroplasia stage (e.g., proliferation and migration of fibroblasts). Finally, the loss of proper eicosanoid response translated into an abnormal migration pattern for the fibroblasts isolated from CERK−/−. As the proper migration of fibroblasts is one of the necessary steps of wound healing, these studies demonstrate a novel requirement for the CERK-derived C1P in the proper healing response of wounds.

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Section: Discussionmentioning

confidence: 99%

Ceramide kinase is required for a normal eicosanoid response and the subsequent orderly migration of fibroblasts

Wijesinghe

Brentnall

Mietla

et al. 2014

Journal of Lipid Research

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“…Normal human epidermal keratinocytes (NHEKs) are usually cultured on 3T3 mouse feeder-layer cells [1]. Several biomaterials have been developed from other animals or humans, but they carry the risk of infection from prions and viruses [2,3].…”

Section: Introductionmentioning

confidence: 99%

Effects of sulfated hyaluronan on keratinocyte differentiation and Wnt and Notch gene expression

2007

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“…In these experimental settings, fibroblasts stop proliferating but remain viable and functional following MMC treatment. 16,17 In a second …”

Section: Min6 Monolayer Cell Viability Following Mitomycin C (Mmc) Trmentioning

confidence: 99%

Down-regulation of proliferation does not affect the secretory function of transformed β-cell lines regardless of their anatomical configuration

Reers

Hauge-Evans

Morgan

et al. 2011

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Mitomycin C-Treated 3T3 Fibroblasts Used as Feeder Layers for Human Keratinocyte Culture Retain the Capacity to Generate Eicosanoids

Cited by 29 publications

References 7 publications

Ceramide kinase is required for a normal eicosanoid response and the subsequent orderly migration of fibroblasts

Ceramide kinase is required for a normal eicosanoid response and the subsequent orderly migration of fibroblasts

Effects of sulfated hyaluronan on keratinocyte differentiation and Wnt and Notch gene expression

Down-regulation of proliferation does not affect the secretory function of transformed β-cell lines regardless of their anatomical configuration

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