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Establishing a High Throughput Epidermal Spheroid Culture System to Model Keratinocyte Stem Cell Plasticity
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JoVE Journal Biologia dello sviluppo
Establishing a High Throughput Epidermal Spheroid Culture System to Model Keratinocyte Stem Cell Plasticity

Establishing a High Throughput Epidermal Spheroid Culture System to Model Keratinocyte Stem Cell Plasticity

DOI:
10.3791/62182-v

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10:03 min

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January 30, 2021

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, , , , ,
1Department of Pathology, Microbiology and Immunology,University of South Carolina School of Medicine, 2Department of Dermatology,Brigham and Women’s Hospital, Harvard Medical School, 3Department of Biochemistry and Molecular Biology,University of Maryland School of Medicine Baltimore, 4Department of Pathology,Albert Einstein College of Medicine, 5Department of Pediatrics,Vanderbilt University Medical Center, 6Department of Drug Discovery and Biomedical Sciences, College of Pharmacy,University of South Carolina

Capitoli

  • 00:04Introduction
  • 00:27Isolation and Culture of Human Keratinocytes from Neonatal Foreskin Tissue
  • 02:37Generating Skin Epidermosphere Cultures in Vitro
  • 05:02Epidermal Spheroid Re-Plating Assay
  • 05:42Characterization of Spheroid-Derived (SD) Sub-Populations by FACS
  • 08:06Results: Characterization of Epidermal Spheroid Formation and Spheroid-Derived NHKs
  • 09:30Conclusion

Summary

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Here we describe a protocol for the systematic cultivation of epidermal spheroids in 3D suspension culture. This protocol has wide-ranging applications for use in a variety of epithelial tissue types and for the modeling of several human diseases and conditions.

Tags

Keywords: Epidermal SpheroidKeratinocyte Stem CellHigh Throughput CultureNeonatal ForeskinDispaseTrypsin-EDTAKSFM-SCM MediumAgarose96-well Plate

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