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Journal of Lipid Research, Vol 33, 1551-1561, Copyright © 1992 by Lipid Research, Inc.
SC Rumsey, NF Galeano, Y Arad and RJ Deckelbaum
The ability to preserve low density lipoprotein (LDL) preparations frozen
for weeks and months without changes in structure or biological properties
is of potential use in long-term comparative studies of LDL. We demonstrate
that freeze-thawing of LDL causes marked alterations in its structure and
biological behavior, and that such changes can be prevented by the addition
of sucrose to the LDL solution prior to freezing. Freezing LDL at -70
degrees C in the absence of sucrose resulted in aggregation and fusion of
particles as measured by electron microscopy, spectrophotometric
absorption, and column gel filtration. This was associated with increased
binding affinity of monoclonal antibodies at epitopes distant from the
receptor binding region. Functional changes induced by freezing included 3-
to 10-fold increases in binding at 4 degrees C and 37 degrees C, and uptake
of LDL in fibroblasts, attributable mainly to increases in nonspecific
binding processes. Cryopreservation of LDL in 10% sucrose (w/v) completely
prevented the structural and functional changes incurred after short- term
freezing, and LDL cryopreserved in sucrose for as long as 18 months
displayed cell binding, uptake, and degradation very similar to that of
freshly obtained LDL.
ARTICLES
Cryopreservation with sucrose maintains normal physical and biological properties of human plasma low density lipoproteins
Department of Pediatrics, Columbia University, New York, NY 10032.
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