Freezing induces artificial cleavage of apoptosis-related proteins in human bone marrow cells.
J Immunol Methods
; 245(1-2): 91-4, 2000 Nov 01.
Article
em En
| MEDLINE
| ID: mdl-11042286
ABSTRACT
The aim of this study was to investigate whether freeze-thawing of freshly isolated human mononuclear bone marrow cells (MNC) influences the integrity of apoptosis-related proteins as determined by immunoblot analyses. Our results show that bone marrow is more sensitive to this process than either myelomonocytoid leukemic P39 or Jurkat T-lymphocyte cell lines. Specifically, bone marrow cells displayed a high level of intrinsic proteolytic activity in response to a single freeze-thaw cycle, which led to the cleavage of various proteins involved in apoptosis cell signaling. This effect was completely blocked by the inclusion of broad-spectrum protease inhibitors in the freezing medium and subsequently thawing the cells on ice. Since differences in the freezing conditions (-80 degrees C vs. liquid nitrogen) did not alter the proteins of interest, we suggest that the thawing process is the critical point when proteolytic enzyme activity is elevated.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Células da Medula Óssea
/
Proteínas
/
Criopreservação
/
Apoptose
Limite:
Humans
Idioma:
En
Revista:
J Immunol Methods
Ano de publicação:
2000
Tipo de documento:
Article
País de afiliação:
Suécia