The anti-cell death FNK protein protects cells from death induced by freezing and thawing.
The FNK protein, constructed from anti-apoptotic Bcl-xL with enhanced activity, was fused with the protein transduction domain (PTD) of the HIV/Tat protein to mediate the delivery of FNK into cells. The fusion protein PTD-FNK was introduced into chondrocytes in isolated articular cartilage-bone sections, cultured neurons, and isolated bone marrow mononuclear cells to evaluate its ability to prevent cell death induced by freezing and thawing. PTD-FNK protected the cells from freeze-thaw damage in a concentration-dependent manner. Addition of PTD-FNK with conventional cryoprotectants (dimethyl sulfoxide and hydroxyethyl starch) increased surviving cell numbers around 2-fold compared with controls treated only with the cryoprotectants. Notably, PTD-FNK allowed CD34+ cells among bone marrow mononuclear cells to survive more efficiently (12-fold more than the control cells) from two successive freeze-thaw cycles. Thus, PTD-FNK prevented cell death induced by freezing and thawing, suggesting that it provides for the successful cryopreservation of biological materials.
Sudo
Kentaro
K
Department of Biochemistry and Cell Biology, Institute of Development and Aging Sciences, Graduate School of Medicine, Nippon Medical School, Kawasaki, Kanagawa 211-8533, Japan.
Asoh
Sadamitsu
S
Ohsawa
Ikuroh
I
Ozaki
Daiya
D
Yamagata
Kumi
K
Ito
Hiromoto
H
Ohta
Shigeo
S
D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D002356:Cartilage / Q000166:cytology / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D019610:Cytoprotection / Q000187:drug effects*; D005615:Freezing; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects; D009474:Neurons / Q000166:cytology / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000494:pharmacology
D018952:Antigens, CD34; D011994:Recombinant Proteins; D017346:Protein Serine-Threonine Kinases