Evaluation of the viability of cultured corneal endothelial cells by quantitative electron probe X-ray microanalysis.

Abstract:

:Construction of artificial organs and tissues by tissue engineering is strongly dependent on the availability of viable cells. For that reason, the viability and the physiological status of cells kept in culture must be evaluated before the cells can be used for clinical purposes. In this work, we determined the viability of isolated rabbit corneal endothelial cells by trypan blue staining and quantitative electron probe X-ray microanalysis. Our results showed that the ionic content of potassium in cultured corneal endothelial cells tended to rise initially, but significantly decreased in cells in the fifth (and final) subculture, especially in comparison to cells in the fourth subculture (P < 0.001). However, the concentration of sulfur was higher in the fifth subculture than in the fourth subculture (P < 0.001), with a nonsignificant increase in sodium in the fifth subculture (P = 0.031). These data imply a remarkable decrease in the K/Na ratio from the fourth to the fifth subculture. Our microanalytical results, along with the morphological differences between cells in the last two subcultures, are compatible with an early phase of the preapoptotic process in the fifth subculture, and suggest that cells of the first four subcultures would be better candidates for tissue engineering.

journal_name

J Cell Physiol

authors

Alaminos M,Sanchez-Quevedo MC,Muñoz-Avila JI,García JM,Crespo PV,González-Andrades M,Campos A

doi

10.1002/jcp.20976

subject

Has Abstract

pub_date

2007-06-01 00:00:00

pages

692-8

issue

3

eissn

0021-9541

issn

1097-4652

journal_volume

211

pub_type

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