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Isolation of Papillary and Reticular Fibroblasts from Human Skin by Fluorescence-activated Cell Sorting
JoVE 杂志
遗传学
This content is Free Access.
JoVE 杂志 遗传学
Isolation of Papillary and Reticular Fibroblasts from Human Skin by Fluorescence-activated Cell Sorting
DOI:

09:14 min

May 07, 2019

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Chapters

  • 00:04Title
  • 01:00Sectioning of Human Skin Dermis into Papillary and Reticular Layers
  • 02:38Separation and Enzymatic Digestion of Epidermis and Dermis
  • 03:52Preparation of Single Cell Suspension and Erythrocyte Lysis
  • 05:58Culturing of Fibroblasts and Adipogenesis Assay
  • 07:04Results: FAP+CD90-, FAP+CD90+ and FAP-CD90+ Fibroblasts Differ in Dermal Localization, Gene Expression and Functionality
  • 08:44Conclusion

Summary

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This manuscript describes a FACS-based protocol for isolation of papillary and reticular fibroblasts from human skin. It circumvents in vitro culture which was inevitable with the commonly used isolation protocol via explant cultures. The emanating fibroblast subsets are functionally distinct and display differential gene expression and localization within the dermis.

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