Document Detail

Procoagulant and profibrinolytic activities of cryopreserved human monocytes.
MedLine Citation:
PMID:  7871496     Owner:  NLM     Status:  MEDLINE    
The purpose of this study was to compare the ability of fresh and cryopreserved mononuclear cells to generate thrombin, induce fibrin formation and finally resolve the fibrin formed, when exposed to plasma. Peripheral blood mononuclear cells (PBM) from 4 donors were collected by gradient centrifugation on Lymfoprep, and cryopreserved in fetal calf serum and 10% dimethyl sulfoxide. Viability was tested by exclusion of trypan blue, as well as green/red fluorescence of fluorescein-diacetate and ethidium bromide (FDA/EB). Fresh and frozen-thawed cells were seeded, stimulated with lipopolysaccharide(LPS), and exposed to a standard heparinized overlay plasma. Plasma was harvested at intervals (0-7 days). Thrombin generation and fibrin formation were measured by quantification of prothrombin fragment (F1 + 2) and fibrinopeptide A (FPA) and the fibrinolytic capacity of the cells as the amount of fibrin (ogen) degradation products (FbDP and FgDP). Recovery of cells after thawing was about 80%, and the viability of fresh and cryopreserved PBM was > 95%. Compared to fresh, frozen cells fully retained their capability of Tissue Factor synthesis, leading to prothombinase activity (F1 + 2) and fibrin formation (FPA). In contrast, the fibrinolytic capacity of frozen-thawed cells were significantly reduced. As expected there were significant variations between the donors in all the parameters measured. We conclude that cryopreservation of human blood mononuclear cells is possible with maintainance of the potential of the cells to mediate coagulation in plasma upon LPS stimulation, whereas the fibrin resolving capacity apparently is reduced by the preservation procedure.
L T Osnes; A B Westvik; P Kierulf
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Thrombosis research     Volume:  76     ISSN:  0049-3848     ISO Abbreviation:  Thromb. Res.     Publication Date:  1994 Nov 
Date Detail:
Created Date:  1995-03-28     Completed Date:  1995-03-28     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0326377     Medline TA:  Thromb Res     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  373-83     Citation Subset:  IM    
Dept. of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway.
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MeSH Terms
Blood Coagulation*
Blood Preservation*
Fibrin / biosynthesis
Fibrin Fibrinogen Degradation Products / analysis
Fibrinopeptide A / analysis
Lipopolysaccharides / pharmacology
Monocytes / drug effects,  physiology*
Peptide Fragments / analysis
Prothrombin / analysis
Thrombin / biosynthesis
Reg. No./Substance:
0/Fibrin Fibrinogen Degradation Products; 0/Lipopolysaccharides; 0/Peptide Fragments; 0/prothrombin fragment 1.2; 25422-31-5/Fibrinopeptide A; 9001-26-7/Prothrombin; 9001-31-4/Fibrin; EC

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