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Seeding of Vascular Grafts with Adipose Derived Adult Stem (ADAS) Cells

用脂肪来源的成体干细胞 (ADAS) 接种血管移植物

基本信息

批准号：
18591418
负责人：
ONOHARA Toshihiro
金额：
$ 2.18万
依托单位：
Department of Clinical Research, National Hospital Organization Kyushu Medical Center (2007)Kyushu University (2006)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591418/
关键词：
Vascular Surgery Regenerative Medicine Artificial Organs

项目摘要

The long-term patency for infrapopliteal bypasses is not favorable in the case using a synthetic vascular graft. The main cause of the synthetic graft failure is that the pseudo-intima of the vascular graft is not covered with regenerated endothelial cells, which results to graft thrombosis in the early stage, or intirnal hyperplasia of the anasotmoses in the late stage. For endothelialization of the graft intima, seeding vascular grafts with endothelial cells had been developed. However, mature endothelial cells have a low proliferative capacity. Thus, seeding vascular grafts with endothelial cells has not been used in the usual clinical settings. Adipose錨erived adult stem (ADAS) cell is a somatic stem cell including fat tissue. ADAS cells are pluripotent cells to differenciate mesenchmal cells, like as fat, bone, muscle, cartilage, and abundantly exist in tissue. Recently, it was reported that ADAS cells can be diffrenciated to endothelial cells. Therefore, endothelialized ADAS culls may be an ideal cell source for a small caliber seeding synthetic graft for infrapopliteal bypass. The purpose of the present study was to develop a seeding vascular graft with ADAS cells. The tissue fragments, which were aspirated from rat subcutaneous adipose tissue, were incubated with cellagenase, and the digest was centrifuged, thereby separating the floating population of mature adipocytes from ADAS cells. Passage culture of ADAS cells was done, and we confirmed that ADAS cells had a high proliferative capacity until 5^<th> passage. After 2 or more passages in cultutu, the expression of endothelial immunophenotype has been investigated in the surface of ADAS cells. Also, we are measuring nitric oxide or prostacyclin production from ADAS cells.

在使用人造血管移植物的情况下，网下搭桥术的长期通畅性是不利的。人工血管移植失败的主要原因是移植血管的假性内膜未被再生的内皮细胞覆盖，导致移植血管早期血栓形成，或晚期移植物原位增生。为了使移植物内膜内皮化，用内皮细胞种植血管移植物已被开发出来。然而，成熟的内皮细胞增殖能力较低。因此，用内皮细胞种植血管移植物并没有在通常的临床环境中使用。脂肪干细胞是一种包括脂肪组织在内的体细胞干细胞，其来源为脂肪细胞(Adpose錨-rive Adminar Stem，ADs)。Adas细胞是分化为间充质细胞的多能细胞，如脂肪、骨、肌肉、软骨等，广泛存在于组织中。最近，有报道称ADAS细胞可以分化为内皮细胞。因此，内皮化的ADAS取材可能是一种理想的小口径种植合成移植物的细胞来源。本研究的目的是建立一种具有ADAS细胞的种子血管移植物。从大鼠皮下脂肪组织中提取组织碎片，用纤维素酶孵育，消化后离心，从而将成熟脂肪细胞的漂浮群体与ADAS细胞分离。对ADAS细胞进行传代培养，证实ADAS细胞在传至第5代时仍具有较高的增殖能力。培养2代或2代以上的ADAS细胞，观察其表面内皮免疫表型的表达。此外，我们还在测量ADAS细胞产生的一氧化氮或前列环素。