Biochemical functionality and recovery of hepatocytes after deep freezing storage |
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Authors: | M Jose Comez-L Pilar Lopez Jose V Castell |
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Institution: | (1) Centro de Investigacion, Hospital La Fe, Ministerio de Sanidad y Consumo, Avda de Campanar 21, Valencia 9, Spain |
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Abstract: | Summary The present study was undertaken to define the conditions for optimal cryopreservation of hepatocytes. Two different freezing
procedures were analyzed: a slow freezing rate (SFR) (−2° C/min down to −30°C and then quick freezing to −196° C) and a fast
freezing rate (FFR) (direct freezing of tubes to −196° C: −39° C/min). Cells were frozen in fetal bovine serum containing
10% Dimethyl sulfoxide (DMSO). After rapid thawing at 37° C, followed by dilution and removal of the cryoprotectant, cells
were plated and several parameters were followed as criteria for optimal cryopreservation of cells. The FFR cells showed no
apparent ultrastructural damage after 24 h of culture. Plating efficiency and spreading were similar as controls. Gluconeogenesis
from pyruvate and fructose, tyrosine amino transferase induction by glucagon and dexamethasone, urea production, and plasma
protein synthesis of FFR cells were similar to those found in control cultures. The FFR procedure, in comparison to the SFR
method, seemed to render the best preserved hepatocytes.
The financial support for this work was from Fondo de Investigaciones Sanitarias de la Seguridad Social, Grants 41/82 and
48/82. |
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Keywords: | cryopreservation hepatocytes primary culture liver cells freezing-thawing gluconeogenesis |
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