Combined PI3K/Akt and Smad2 activation promotes corneal endothelial cell proliferation

Alfonso L. Sabater, Enrique J. Andreu, María García-Guzmán, Tania López, Gloria Abizanda, Victor L. Perez, Javier Moreno-Montañés, Felipe Prósper

Research output: Contribution to journalArticlepeer-review

10 Scopus citations


PURPOSE. The purpose of this study was to develop a culture method for expansion of corneal endothelial cells (CEC) based on the combined activation of PI3K/Akt and Smad2. METHODS. Morphology, proliferation, and migration of cultured rabbit and nonhuman primate CEC were examined in the presence of the PI3K/Akt activators IGF-1 and heregulin beta in combination with the Smad2 activator activin A. Phenotypic characterization of CEC was performed at the RNA and protein levels. Cell pump function and transepithelial electric resistance were used for in vitro functional assessment of CEC. Finally, ex vivo-expanded rabbit CEC were transplanted into a model of endothelial damage in rabbit corneas. RESULTS. Treatment of rabbit and nonhuman primate CEC in vitro with IGF-1, heregulin beta, and activin A induced an upregulation of PI3K/Akt and Smad2 signaling pathways and an increase in proliferation and migration of CEC expressing ZO-1, connexin-43, and Naþ/Kþ- ATPase. Cell pump function evaluation revealed the complete functionality of cultured CEC. Injection of rabbit CEC successfully produced recovery of normal corneal thickness in a rabbit model of endothelial dysfunction. CONCLUSIONS. We demonstrated that the combined activation of PI3K/Akt and Smad2 results in in vitro expansion of phenotypic and functional CEC. Expanded cells were able to contribute to restoration of corneal endothelium in a rabbit model. These findings may represent a new therapeutic approach for treating corneal endothelial diseases.

Original languageEnglish (US)
Pages (from-to)745-754
Number of pages10
JournalInvestigative Ophthalmology and Visual Science
Issue number2
StatePublished - Feb 1 2017


  • Corneal endothelial cell proliferation
  • Corneal endothelial cell transplantation
  • Corneal endothelial regeneration

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience


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