Nakayama S, Ban T, Okamoto Y
Department of Cardiovascular Surgery, Faculty of Medicine, Kyoto University, Japan.
Nihon Kyobu Geka Gakkai Zasshi. 1993 Aug;41(8):1357-62.
The purpose of this study is to clarify whether serum (fetal bovine serum or human serum) is necessary or not for cryopreservation of aortic valve allografts. The protective effects of fetal bovine serum compared with human serum were evaluated by means of quantitative assessment of fibroblast viability. Porcine aortic valves were excised and rinsed immediately after death, followed by treatment with low concentration antibiotics. Valves were gradually frozen at a control-rate of -1 degree C/min and then stored in liquid nitrogen vapor-phase. The samples were classified into three groups by nutrient medium as follows. Group A (n = 5); Dulbecco's Modified Eagle Medium (DMEM), Group B (n = 5); DMEM containing 10% fetal bovine serum, Group C (n = 5); DMEM containing 20% human serum. At 1 week, 1 month, and 3 months after initiation of storage, the valves were thawed rapidly and examined for fibroblast viability, which was assessed quantitatively by means of autoradiography with tritiated proline. After 1 week storage, the fibroblast viability rate reduced from 98% to 80.7% in group A, 79.4% in group B and 79.2% in group C respectively, but there were no significant differences among three groups. The viability rate decreased with the passage of time, and remained around 72% in all groups after 3 months storage. The histological examination did not show any changes in the structure of the valves in all samples up to 3 months of storage. The results demonstrate that the fibroblasts are still viable after 3 months storage by cryopreservation. The addition of fetal bovine serum or human serum does not improve fibroblast viability.(ABSTRACT TRUNCATED AT 250 WORDS)
本研究的目的是阐明血清(胎牛血清或人血清)对于同种异体主动脉瓣冷冻保存是否必要。通过对成纤维细胞活力的定量评估,比较了胎牛血清与人血清的保护作用。猪主动脉瓣在猪死后立即切除并冲洗,然后用低浓度抗生素处理。瓣膜以-1℃/分钟的控制速率逐渐冷冻,然后储存在液氮气相中。样本根据营养培养基分为三组,如下所示。A组(n = 5);杜氏改良 Eagle 培养基(DMEM),B组(n = 5);含10%胎牛血清的DMEM,C组(n = 5);含20%人血清的DMEM。在储存开始后的1周、1个月和3个月,瓣膜迅速解冻并检测成纤维细胞活力,通过用氚标记的脯氨酸进行放射自显影定量评估。储存1周后,A组的成纤维细胞活力率分别从98%降至80.7%,B组为79.4%,C组为79.2%,但三组之间无显著差异。活力率随时间下降,储存3个月后所有组均保持在72%左右。组织学检查显示,在储存长达3个月的所有样本中,瓣膜结构均未出现任何变化。结果表明,通过冷冻保存3个月后成纤维细胞仍具有活力。添加胎牛血清或人血清并不能提高成纤维细胞活力。(摘要截断于250字)