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Primary mouse keratinocyte culture
Annalisa Pirrone1, Barbara Hager, Philip Fleckman
1Department of Medicine, University of Seattle, WA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|October 27, 2004
Summary
This study details a new method for culturing mouse epidermal keratinocytes, enabling long-term cell growth and analysis. This technique is crucial for research involving transgenic or knockout mice with skin conditions.
Area of Science:
- Dermatology
- Cell Biology
- Developmental Biology
Background:
- Mouse epidermal keratinocytes are challenging to culture in vitro.
- Existing methods limit long-term studies and analysis of genetically modified mice.
Purpose of the Study:
- To present a robust protocol for isolating and culturing mouse epidermal keratinocytes.
- To facilitate long-term cell culture and analysis, particularly for transgenic and knockout mouse models.
- To provide detailed methods for inducing differentiation, shipping skins, and cryopreserving cells.
Main Methods:
- Isolation of epidermal keratinocytes from single newborn mouse pups.
- Utilization of supplemented fibroblast-conditioned medium and collagen IV-coated dishes.
- Establishment of protocols for subculturing, inducing differentiation markers, shipping skins, and cryopreservation.
Main Results:
- Successful long-term culture and multiple passages of mouse epidermal keratinocytes.
- Demonstrated induction of late-stage epidermal differentiation markers in vitro.
- High cell viability achieved after cryopreservation and storage in liquid nitrogen.
Conclusions:
- The presented method overcomes challenges in culturing mouse epidermal keratinocytes.
- This protocol is valuable for researchers studying epidermal development and defects in transgenic or knockout mice.
- The techniques support diverse research applications, including remote sample processing and long-term cell banking.