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KMID : 0352519940310030449
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Korea Univercity Medical Journal
1994 Volume.31 No. 3 p.449 ~ p.460
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Viability of the Endothelial cells using Flow Cytometry in Aortic and Pulmonary
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Abstract
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Human allograft valves have been widely applied for the treatment of acquired and congenital cardiac diseases. One factor that some investigators consider to be important in determining allograft durability is the degree of cellular
viability. Allografts comprise a heterogenous population of cell and determination of graft viability logically requires
understanding viability of those specific populations.
This study was performed to quantify the viability of endothelial cells of allograft cardiac valves after 4。C fresh preservation and -196。C cryopreservation for 14 ays using metabolic assay technique with 3H-glycine and flow
cytometric
technique. To evaluate endothelial cell viability using Flow Cytometry, valved conduit were subjected to collagenase digestion. The resulting cell suspension was labeled with Griffonia simplicifolia agglutinin-fluorescein isothiocyanate
(GSA-FITC)
a marker for vascular endothelial cells. The cells were then incubated with propidium iodide (PI), which is excluded by viable cells. Flow cytometry evaluated endothelial cell viability by determination of percentage of GSA-FITC-positive
cells
that were negative for PI.
The metabolic assay shows that the viability of endothelial cells was 49% after cryopreservation and 17% after fresh preservation for 14 days.
The flow cytometric analysis shows that viability of endothelial cells was 39% after
cryopreservation and 41% after fresh preservation for 14days.
Flow cytometry is fast and precise quantitative method for evaluating the viability of
endothelial cells.
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