| Effect of Cryopreservation of Human Heart Cells on Cell Proliferation |
(Division of Cardiovascular Surgery, Department of Surgery (Omori), School of Medicine, Faculty of Medicine and Department of Biomolecular Science, Faculty of Science*, Toho University, Tokyo, Japan)
| Hiroki Yokomuro |
Noritsugu Shiono |
Tsukasa Ozawa |
| Takeshirou Fujii |
Muneyasu Kawasaki |
Yoshinori Watanabe |
| Katsunori Yoshihara |
Nobuya Koyama |
Mitsumasa Okada* |
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Preservation is essential for successful cell transplantation. 1) Control group (n13); Cells isolated from human right atrial tissues were cultured for 15 days. 2) Cell-cryopreservation (C.P.) group (n23), Tissue-C.P. group (n29); Human heart cells and minced tissues were cryopreserved in freezing medium containing 70% IMDM, 20% FBS, and 10% DMSO at a rate of|1/min. to |80 by a programmed freezer and stored in liquid nitrogen (|196) for 1 week. After cryopreservation, the tissues and cells were thawed rapidly at 37. The cells, cryopreserved cells and cells isolated from cryopreserved tissues were cultured as passage 1, 2, and 3 for 15 days each. Cell proliferation was compared with a control group by determining growth curves, and 2-day proliferation rates. A growth factor, biochemical features and cell cycle were measured pre and post-cryopreservation. The cryopreserved group proliferated much more than the control group within 15 days at passage 1, 2, and 3 (1.7, 2.1, and 3.1 times, p0.0001) respectively. The 2-day proliferation rates of cryopreservation group were higher than the control group in 15 days (p0.05). The bFGF release after cryopreservation was on average 46.8 and 6.8 times greater than before cryopreservation for the Cell-C.P. and Tissue-C.P. groups, respectively. The TGF-ภ1 release was also accelerated by cryopreservation (Cell-C.P. group: 1.78 times, Tissue-C.P. group: 1.45 times in average) after cryopreservation. The cell cycle of human heart cells shifted to G2{M from the G1{G0 period by cryopreservation. Human atrial tissues and cells can be cultured and cryopreserved. The cryopreserved cells and cells isolated from cryopreserved tissue proliferate much more than non-cryopreserved cells at all cell ages. Cryopreservation enables human tissues and cells to proliferate more because of the greater release of growth factors and changing cell cycle.
@Jpn. J. Cardiovasc. Surg. 35: 14-20 (2006) |
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