Ruddle A C, George S J, Armitage W J, Alexander E L, Mitchell D C
Department of Vascular Surgery, Southmead Hospital, Bristol, UK.
Eur J Vasc Endovasc Surg. 2000 Mar;19(3):233-7. doi: 10.1053/ejvs.1999.0989.
we assessed the effects of cryopreservation on smooth-muscle cell injury in human vein.
long saphenous vein was collected during surgery and cryopreserved. Smooth-muscle cell damage was assessed after thawing by in situ detection of fragmented DNA. The presence of cryoprotectant (10% dimethyl sulphoxide, DMSO), cooling and warming rates, and the rate of cryoprotectant removal after thawing were examined.
control veins exhibited damage in 8.5% (95% confidence interval (CI) 4.7 to 13.4%,n=13) of smooth-muscle cells compared with 27.7% (95% CI 23.2 to 32.4%, n=115) in vein frozen in 10% DMSO (p=0.001). In the presence of DMSO, damage to smooth-muscle cells was independent of the rates of cooling (p=0.72) and warming (p=0.45). The rate of dilution to remove the cryoprotectant after thawing also had no effect on cell damage (p=0.64). In the absence of cryoprotectant, cell damage was doubled to approximately 50% by slow rather than rapid warming (p=0.01).
cooling rate, and the presence of a cryoprotectant, has little effect on smooth-muscle damage, provided that the tissue is warmed rapidly. Slow warming, in the absence of DMSO, causes substantial damage. These results suggest that simplified methods of vein cryopreservation are feasible.
我们评估了冷冻保存对人静脉平滑肌细胞损伤的影响。
在手术过程中采集大隐静脉并进行冷冻保存。解冻后通过原位检测DNA片段评估平滑肌细胞损伤。检测了冷冻保护剂(10%二甲亚砜,DMSO)的存在、降温与升温速率以及解冻后冷冻保护剂的去除速率。
与在10%DMSO中冷冻的静脉相比,对照静脉中8.5%(95%置信区间(CI)4.7至13.4%,n = 13)的平滑肌细胞出现损伤,而在10%DMSO中冷冻的静脉中这一比例为27.7%(95%CI 23.2至32.4%,n = 115)(p = 0.001)。在存在DMSO的情况下,平滑肌细胞损伤与降温速率(p = 0.72)和升温速率(p = 0.45)无关。解冻后去除冷冻保护剂的稀释速率对细胞损伤也没有影响(p = 0.64)。在没有冷冻保护剂的情况下,缓慢升温而非快速升温使细胞损伤增加一倍,达到约50%(p = 0.01)。
只要组织快速升温,降温速率和冷冻保护剂的存在对平滑肌损伤影响很小。在没有DMSO的情况下缓慢升温会导致严重损伤。这些结果表明简化的静脉冷冻保存方法是可行的。