Orita H, Fukasawa M, Inui K, Hirooka S, Uchino H, Fukui K, Kohi M, Washio M
Second Department of Surgery, Yamagata University School of Medicine, Japan.
Jpn Heart J. 1994 Mar;35(2):225-35. doi: 10.1536/ihj.35.225.
In this study, we evaluated cardiac myocyte viability and function under hypothermic conditions using three types of storage solutions; saline solution (SS), Euro-Collins solution (ECS) and MCDB 107 medium (MM). Cardiac myocytes were isolated from neonatal rat ventricles by collagenase dispersion and cultured for 4 days with MCDB 107 medium. A total of 12.5 x 10(5) myocytes/culture dish were used and the myocytes were incubated at 4 degrees C for 6, 12, 18 and 24 hrs in the various storage solutions. After each incubation time, CPK and LDH were measured in the storage solutions. The myocytes were then cultured in MCDB 107 medium and incubated for 24 hrs at 37 degrees C to evaluate the recovery of the myocyte beating rate. In group MM (n = 7), the recovery ratio of the myocyte beating rate was 99.2 percent of control (beating rate prior to hypothermic incubation) at 6 hours, 104.6% at 12 hrs and 44.8% at 24 hrs. Groups SS and ECS (n = 7 each) had significantly lower recovery ratios than the MM group (at 6 hrs: 74.3, 34.0; at 12 hrs: 61.0, 32.2; at 24 hrs: 0.0, 0.0 percent of control, respectively). Release of CPK and LDH in the MM group gradually increased and at 24 hrs was 28.6 IU/l and 93.6 IU/l, respectively. However, the SS group had significantly increased CPK and LDH values at 24 hrs (CPK: 66.9, LDH: 164.2). The ECS group showed the greatest increase in both markers (CPK: 317.5, LDH: 421.2). In summary, saline solution showed a beneficial effect on recovery of myocyte viability at 12 hours compared to Euro-Collins solution, however, MCDB 107 medium had the best overall protective effect on cultured myocytes. Accordingly, alternate hypothermic storage solutions, such as cell-culture medium, may have protective characteristics that are suitable for cardiac preservation.
在本研究中，我们使用三种保存溶液评估了低温条件下心肌细胞的活力和功能；这三种溶液分别是盐溶液（SS）、欧洲柯林斯溶液（ECS）和MCDB 107培养基（MM）。通过胶原酶分散法从新生大鼠心室中分离出心肌细胞，并用MCDB 107培养基培养4天。每个培养皿中使用总共12.5×10⁵个心肌细胞，并将这些心肌细胞在4℃下于各种保存溶液中孵育6、12、18和24小时。在每次孵育时间后，测量保存溶液中的肌酸磷酸激酶（CPK）和乳酸脱氢酶（LDH）。然后将心肌细胞置于MCDB 107培养基中培养，并在37℃下孵育24小时，以评估心肌细胞搏动率的恢复情况。在MM组（n = 7）中，心肌细胞搏动率的恢复率在6小时时为对照组（低温孵育前的搏动率）的99.2%，12小时时为104.6%，24小时时为44.8%。SS组和ECS组（每组n = 7）的恢复率显著低于MM组（6小时时分别为74.3、34.0；12小时时分别为61.0、32.2；24小时时分别为对照组的0.0、0.0%）。MM组中CPK和LDH的释放逐渐增加，24小时时分别为28.6 IU/L和93.6 IU/L。然而，SS组在24小时时CPK和LDH值显著升高（CPK：66.9，LDH：164.2）。ECS组的这两种标志物升高幅度最大（CPK：317.5，LDH：421.2）。总之，与欧洲柯林斯溶液相比，盐溶液在12小时时对心肌细胞活力的恢复显示出有益作用，然而，MCDB 107培养基对培养的心肌细胞具有最佳的总体保护作用。因此，诸如细胞培养基之类的替代性低温保存溶液可能具有适合心脏保存的保护特性。
