The anti-cell death FNK protein protects cells from death induced by freezing and thawing |
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Authors: | Sudo Kentaro Asoh Sadamitsu Ohsawa Ikuroh Ozaki Daiya Yamagata Kumi Ito Hiromoto Ohta Shigeo |
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Affiliation: | Department of Biochemistry and Cell Biology, Institute of Development and Aging Sciences, Graduate School of Medicine, Nippon Medical School, Kawasaki, Kanagawa 211-8533, Japan. |
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Abstract: | 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. |
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Keywords: | Bcl-xL Apoptosis Cell death Cryopreservation Cartilage Chondrocyte Neuron Bone marrow mononucleocyte CD34 Protein transduction domain |
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