Culture of human limbal epithelial stem cells on tenon’s fibroblast feeder-layers: A translational approach

Gaia Scafetta, Camilla Siciliano, Giacomo Frati, Elena De Falco

Research output: Contribution to journalArticlepeer-review

Abstract

The coculture technique is the standard method to expand ex vivo limbal stem cells (LSCs) by using inactivated embryonic murine feeder layers (3T3). Although alternative techniques such as amniotic membranes or scaffolds have been proposed, feeder layers are still considered to be the best method, due to their ability to preserve some critical properties of LSCs such as cell growth and viability, stemness phenotype, and clonogenic potential. Furthermore, clinical applications of LSCs cultured on 3T3 have taken place. Nevertheless, for an improved Good Manufacturing Practice (GMP) compliance, the use of human feeder-layers as well as a fine standardization of the process is strictly encouraged. Here, we describe a translational approach in accordance with GMP regulations to culture LSCs onto human Tenon’s fibroblasts (TFs). In this chapter, based on our experience we identify and analyze issues that often are encountered by researchers and discuss solutions to common problems.

Original languageEnglish
Pages (from-to)187-198
Number of pages12
JournalMethods in molecular biology (Clifton, N.J.)
Volume1283
DOIs
Publication statusPublished - 2015

Keywords

  • Cell therapy
  • Feeder layers 3T3
  • Good manufacturing practices (GMPs)
  • Human Tenon’s fibroblasts
  • Limbal stem cells

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Medicine(all)

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