Tahara K, Uchida H, Kawarasaki H, Hasizume K, Kobayashi E
Division of Organ Replacement Research, Center for Molecular Medicine, Jichi Medical School, 3-1-1, Yakushiji, Minamikawachi, Kawachi, Tochigi 332-0498, Japan.
J Pediatr Surg. 2001 Apr;36(4):602-4. doi: 10.1053/jpsu.2001.22295.
If long-term organ cryopreservation can be attained, a significant achievement will have been made to address the problem for donor shortage. Fetal intestine has been known to revascularize naturally without vascular anastmosis. The authors have confirmed previously that the newborn intestine also could develop to maturity in the host omentum. Here, the authors examined whether the cryopreserved newborn intestine could revascularize in the syngeneic combination using the 2 different solutions and whether cryopreservation affect their antigenicity in the allogeneic combination.
Inbred rat strains of LEW (MHC haplotype; RT1(l)) and BN (RT1(n)) were used. LEW newborn intestinal grafts were stored in RPMI-1640 or University of Wisconsin solution with 10% DEMSO (n = 10 in each group). The grafts were placed into a cold (4 degrees C) preservation solution for 30 minutes and then placed into a freezing chamber and cooled to -80 degrees C at -1 degrees C/min after 12 hours quenched to -180 degrees C in liquid nitrogen for longer than 30 days. Then, the cryopreserved grafts under the 2 different solutions were transplanted syngenicaly (LEW to LEW). The cryopreserved BN grafts also were implanted into the LEW omentum pouch. The allotransplantation was received with a 14-day high-dose course of tacrolimus (0.64 mg/kg, intramuscularly). The grafts were evaluated histologically at 4 weeks after transplantation. Fresh newborn intestines implanted in this syngeneic and allogeneic combination were evaluated as each control group.
In the syngeneic combination, more than 90% of the mature intestine were obtained. There was no significant difference among the different solution and the fresh group. However, in the allogeneic combination, both fresh and cryopreserved grafts were histologically poor.
This is the first report showing that long-term cryopreservation was not harmful for neovascularization of newborn intestine. Long-term cryopreservation did not reduce the antigenicity of the newborn intestine. J Pediatr Surg 36:602-604.
如果能够实现长期器官冷冻保存,将在解决供体短缺问题方面取得重大进展。已知胎儿肠道可自然实现血管再生,无需进行血管吻合。作者之前已经证实,新生肠道在宿主网膜中也可发育成熟。在此,作者研究了使用两种不同溶液冷冻保存的新生肠道在同基因组合中是否能够实现血管再生,以及冷冻保存是否会影响其在异基因组合中的抗原性。
使用近交系大鼠LEW(MHC单倍型;RT1(l))和BN(RT1(n))。将LEW新生肠道移植物保存在含有10%二甲基亚砜的RPMI-1640或威斯康星大学溶液中(每组n = 10)。将移植物置于冷(4℃)保存溶液中30分钟,然后放入冷冻室,在12小时后以-1℃/分钟的速度冷却至-80℃,再在液氮中骤冷至-180℃并保存超过30天。然后,将两种不同溶液下冷冻保存的移植物进行同基因移植(LEW到LEW)。冷冻保存的BN移植物也植入LEW网膜袋中。接受14天高剂量他克莫司(0.64mg/kg,肌肉注射)的同种异体移植。在移植后4周对移植物进行组织学评估。将植入这种同基因和异基因组合的新鲜新生肠道作为每个对照组进行评估。
在同基因组合中,获得了超过90%的成熟肠道。不同溶液组与新鲜组之间无显著差异。然而,在异基因组合中,新鲜和冷冻保存的移植物在组织学上均较差。
这是第一份表明长期冷冻保存对新生肠道新生血管形成无害的报告。长期冷冻保存并未降低新生肠道的抗原性。《小儿外科杂志》36:602 - 604。