Clonal analysis of human clonogenic keratinocytes.

Abstract

Regenerative medicine has its roots in harnessing stem cells for permanent restoration of damaged or diseased tissues. The first procedure for the transplantation of epidermal cultures in massive full-thickness burns was established in the 1980s. Since then, epithelial stem cell-based therapies have been further developed in cell and gene therapy protocols aimed at restoring visual acuity in severe ocular burns and treating patients affected by genetic skin diseases, as Epidermolysis Bullosa. The clinical success of these Advanced Therapy Medicinal Products (ATMPs) requires the presence of a defined number of epithelial stem cells in the grafts, detected as holoclone-forming cells. To date, the most trustworthy method to identify and measure holoclones in a culture is the clonal analysis of clonogenic keratinocytes. Here we describe in detail how to perform such a clonal analysis and identify each epidermal clonal type.

• Authors: Elena Enzo, Camilla Cattaneo, Federica Consiglio, Maria Pia Polito, Sergio Bondanza, Michele De Luca
• Journal: Methods in cell biology (Methods Cell Biol) Vol. 170 Pg. 101-116 ( 2022) ISSN: 0091-679X [Print] United States
• PMID: 35811094 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2022 Elsevier Inc. All rights reserved.

Topics

• Cells, Cultured
• Genetic Therapy (methods)
• Humans
• Keratinocytes
• Regenerative Medicine
• Stem Cells