Orita H, Fukasawa M, Hirooka S, Fukui K, Kohi M, Washio M, Sasaki H
Second Department of Surgery, Yamagata University School of Medicine, Japan.
J Surg Res. 1993 Dec;55(6):654-8. doi: 10.1006/jsre.1993.1199.
In this study, we evaluated the functional and biochemical effects of cardiac fibroblasts (CFs) on cardiac myocytes (CMs) incubated under hypothermic conditions. CMs and CFs were isolated from neonatal rat ventricles and cultures of myocytes only or myocytes in coculture with CFs were established. A CM and CF concentration of 2.5 x 10(5) cells/ml was chosen and the optimum proportion of CF was determined to be 33% (ml CM:ml CF) as follows: 6:0 (group C-a), 4:0 (group C-b), 4:2 (group M-F). On the fourth day of culture, cells were incubated at 4 degrees C for 6, 12, 18, 24, 36 and 48 hr. After each incubation, creatine phosphokinase ((PK) and lactate dehydrogenase (LDH) were measured in the medium, and then the cells were cultured for an additional 24 hr at 37 degrees C to evaluate the recovery of the CM contracting rate. In groups C-a and C-b (n = 6 each), the recovery ratio of CM contracting rate decreased at 18 hr (C-a, 59.0; C-b, 55.6% of control; i.e., contracting rate prior to hypothermic incubation), reaching null levels at 48 hr. However, group M-F (n = 6) showed a significantly increased recovery at 18 hr (95.7%, P < 0.05; at 48 hrs, 27.6%). Release of CPK and LDH in groups C-a and C-b increased gradually by 24 hr and showed a marked increase at 48 hr (C-a, 114.3 x 10(-2), 194.2 x 10(-2); C-b, 81.6 x 10(-2), 118.6 x 10(-2) IU/flask, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)
在本研究中，我们评估了心脏成纤维细胞（CFs）对在低温条件下培养的心肌细胞（CMs）的功能和生化影响。从新生大鼠心室中分离出CMs和CFs，并建立了仅培养心肌细胞或心肌细胞与CFs共培养的体系。选择CM和CF的浓度为2.5×10⁵个细胞/ml，并确定CF的最佳比例为33%（CM毫升数:CF毫升数），如下：6:0（C-a组）、4:0（C-b组）、4:2（M-F组）。在培养的第4天，将细胞在4℃下孵育6、12、18、24、36和48小时。每次孵育后，测定培养基中的肌酸磷酸激酶（CPK）和乳酸脱氢酶（LDH），然后将细胞在37℃下再培养24小时以评估CM收缩率的恢复情况。在C-a组和C-b组（每组n = 6）中，CM收缩率的恢复率在18小时时下降（C-a组为对照组的59.0%；C-b组为55.6%，即低温孵育前的收缩率），在48小时时降至零水平。然而，M-F组（n = 6）在18小时时显示出显著增加的恢复率（95.7%，P < 0.05；在48小时时为27.6%）。C-a组和C-b组中CPK和LDH的释放量在24小时时逐渐增加，并在48小时时显著增加（C-a组分别为114.3×10⁻²、194.2×10⁻²；C-b组分别为81.6×10⁻²、118.6×10⁻² IU/培养瓶）。（摘要截断于250字）
