Optimization of Human Limbal Stem Cell Culture by Replating a Single Limbal Explant

Part of the Methods in Molecular Biology book series (MIMB, volume 2145)


Cultured limbal epithelial stem cell transplantation is a clinical procedure used to regenerate the corneal epithelium in patients with limbal stem cell deficiency. The protocols used to expand limbal epithelial cells in vitro need to be optimized, since the scarcity of human ocular tissue donors is limiting the potential use of this procedure. Here, we describe a method to consecutively expand a single human limbal explant. With this method it is possible to obtain up to three limbal epithelial primary cultures from the same explant, thus increasing the efficiency of the in vitro cell culture.

Key words

Cornea Corneal epithelium Corneoscleral samples Limbus Limbal explant Limbal stem cells Limbal primary cultures In vitro expansion 



Financial Support: Ministerio de Economía y Competitividad and Fondo Europeo de Desarrollo Regional, Spain (SAF2015-63594-R MINECO/FEDER, EU). Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN). Instituto de Salud Carlos III, Spain. Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, Spain. 


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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Grupo de Superficie OcularInstituto Universitario de Oftalmobiología Aplicada (IOBA), Universidad de ValladolidValladolidSpain
  2. 2.Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN). Instituto de Salud Carlos IIIMadridSpain
  3. 3.Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, ValladolidSpain

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