Revista Sociedad Colombiana de Oftalmología - Volumen 44 (3) Julio - Septiembre 2011
colágeno que luego de varios sub-cultivos, permitieron obtener una monocapa celular de características morfológicas similares a las del endotelio nativo. Los cambios en el recuento celular fueron medidos mediante microscopía y usando tinción de azul tripán, así mismo se usó para probar viabilidad celular. Resultados: La técnica seleccionada permite obtener una monocapa celular con alta densidad celular y características morfológicas similares a las del endotelio nativo. La técnica seleccionada usa quelantes iónicos y superficies tratadas con componentes de la matriz extracelular. La viabilidad celular con este método es cercana al 100%. Conclusiones: Existen diferencias importantes entre el cultivo de células del endotelio de conejo y el procedimiento realizado para endotelio corneal humano. La obtención de una monocapa de endotelio corneal humano es especialmente significativa considerando la posibilidad de implementar una nueva técnica para el tratamiento de las lesiones del endotelio de la córnea. Palabras clave: Endotelio corneal, cultivo celular, medicina regenerativa.
Methods: Primary cultures of corneal endothelial cells were prepared from rabbit eyes using plastic and glass surfaces with different isolation methods to standardize the culture conditions. The human endothelial cells of donor corneas were detached from Descemet’s membrane and were placed in plates covered with collagen and growth medium. After several passages the changes in density of the corneal endothelium were quantified by microscopy and counting of trypan-blue–stained cells. Results: We searched for the best media conditions and procedures that resulted in high density cell cultures and morphological characteristics similar to native corneal endothelium tissues. The selected technique among those tried consisted of using divalent chelating agents and surfaces treated with components of extracellular matrix such as collagen type IV. With this technique, cellular viability was found to be close to 100%. Conclusions: Major differences have been identified between cultured corneal endothelial cells in human and rabbits. The culture of human corneal endothelial cells is especially important as they present a new approach for the treatment of corneal endothelium deficiency.
Abstract Purpose: Corneal endothelial cells in humans do not replicate to any meaningful extent. Diminishing density of the cell monolayer is a major cause of loss of corneal transparency. This paper reports the successful tissue cultures of corneal endothelial cells in monolayers using peripheral regions of the cornea.
Keywords: corneal endothelial cells, cell culture, regenerative medicine. El endotelio es la capa más interna de la córnea, contribuye al mantenimiento de la transparencia corneal dada su función de barrera y de bomba hidroelectrolítica. En humanos su capacidad de multiplicación
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