Adipose tissue-derived stem cells in a fibrin implant enhance neovascularization in a peritoneal grafting site: a potential way to improve ovarian tissue transplantation
Adipose tissue-derived stem cells in a fibrin implant enhance neovascularization in a peritoneal grafting site: a potential way to improve ovarian tissue transplantation
复制标题
DOI：
10.1093/humrep/dex374
复制
发表时间：
2018-02-01
影响因子：
6.1
通讯作者：
Dolmans, M. M.
中科院分区：
文献类型：
--
作者：
Manavella, D. D.;Cacciottola, L.;Dolmans, M. M.
两种不同浓度的人脂肪组织来源的干细胞（ASC）包埋在纤维蛋白支架内是否有分化成血管的潜力，并有助于卵巢组织移植的腹膜移植部位的血管化？当移植到纤维蛋白基质内的小鼠腹膜时，低浓度和高浓度的人ASC分化成血管，但仅高ASC浓度在移植后14天显著增加人血管面积ASC具有多谱系分化潜力，包括促血管生成特性，并且已用于组织工程以增强移植组织中的血管形成。利用22只严重联合免疫缺陷小鼠建立体内实验模型。总共有16只小鼠（每组8只）腹腔内移植了载有两种不同人ASC浓度（150 000 [L-ASC]或1 500 000 [H-ASC]细胞）和酞菁锂（LiPc）晶体作为氧敏感探针的纤维蛋白支架。六只小鼠移植有仅含有LiPc的空纤维蛋白（EF）植入物，并作为对照。通过电子顺磁共振血氧测定法（EPR）在体内监测植入物中的氧分压（pO（2））水平。通过免疫组织化学（IHC）分析ASC鉴定、增殖以及宿主和人血管形成。所有分析均在移植后第3、7和14天进行。前瞻性实验研究在天主教鲁汶大学妇科研究中心进行。所有材料均用于进行pO（2）测量（EPR血氧测定法）、组织学（苏木精-伊红染色）和IHC（抗人波形蛋白、抗人Ki 67、抗小鼠和人双CD 34）分析。从第7天到第14天，ASC负载的植入物显示出pO（2）水平增加的趋势，而EF植入物中未观察到。加载ASC的植入物显示分化为人CD 34阳性血管。总CD 34阳性内皮面积与EPR血氧测定法获得的pO（2）值相关（r = 0.6506，P = 0.0019）。在H-ASC组中，在移植后14天，与L-ASC组相比，发现更大的人CD 34阳性血管表面积（P < 0.0049）。然而，其他可能的机制参与ASC血管生成的行为仍有待调查。高浓度的ASCs装载在纤维蛋白支架内可以作为一个基板，准备腹膜移植网站超过14天，以提高血管化一旦人类卵巢组织移植。本研究得到了Fonds National de la Recherche Scientifique de Escherque的赠款（FNRS-PDR Convention T.0077.14，T，l，vie授予DDM的资助号7.6515.16 F和授予M.M.D.的资助号5/4/150/5）。[CAA是FRS-FNRS研究助理]）、Fonds Sp、ciaux de Recherche和Fondation St Luc、抗癌基金会以及费列罗家族的捐款。没有一位作者有任何利益冲突需要声明。
Do two different concentrations of human adipose tissue-derived stem cells (ASCs) embedded inside a fibrin scaffold have the potential to differentiate into vessels and aid vascularization in a peritoneal grafting site intended for ovarian tissue transplantation?Human ASCs in low and high concentrations differentiated into vessels when transplanted to mouse peritoneum inside a fibrin matrix, but only high ASC concentrations significantly increased human vessel area 14 days after transplantation.ASCs have multilineage differentiation potential, including proangiogenic properties and have been used in tissue engineering to enhance vascularization in transplanted tissues. Fibrin has been studied and used as an ASC-compatible biomaterial.In vivo experimental model using 22 severe combined immunodeficient mice. In total, 16 mice (eight per group) were intraperitoneally grafted with a fibrin scaffold loaded with two different human ASC concentrations (either 150 000 [L-ASC] or 1 500 000 [H-ASC] cells) and lithium phthalocyanine (LiPc) crystals as oxygen-sensitive probes. Six mice were grafted with an empty fibrin (EF) implant containing only LiPc and served as controls. Levels of partial pressure of oxygen (pO(2)) in implants were monitored in vivo by electron paramagnetic resonance oximetry (EPR). ASC identification, proliferation, and host and human vascularization were analyzed by immunohistochemistry (IHC). All analyses were performed on post-grafting Days 3, 7 and 14.Prospective experimental study conducted at the Gynecology Research Unit, Universit, Catholique de Louvain. All materials were used to perform pO(2) measurements (EPR oximetry), as well as histological (hematoxylin-eosin staining) and IHC (anti-human vimentin, anti-human Ki67, anti-mouse and human double CD34) analyses.A significant increase in pO(2) in implants was observed in all groups between Days 3 and 7 (P < 0.001). ASC-loaded implants displayed a tendency towards increased pO(2) levels from Days 7 to 14, not observed in EF implants. ASC-loaded implants showed differentiation into human CD34-positive vessels. Total CD34-positive endothelial area was correlated to pO(2) values obtained by EPR oximetry (r = 0.6506, P = 0.0019). In the H-ASC group, a greater human CD34-positive vascular surface area was found compared to the L-ASC group 14 days after transplantation (P < 0.0049).N/A.As demonstrated by our results, ASCs transplanted inside a fibrin matrix can differentiate into CD34-positive human vessels. However, other possible mechanisms involved in ASC angiogenic behavior remain to be investigated.High concentrations of ASCs loaded inside a fibrin scaffold could serve as a substrate to prepare a peritoneal grafting site over 14 days, in order to enhance vascularization once human ovarian tissue is grafted. Our proposed preparation of the grafting site would not only benefit ovarian tissue transplantation, but also other experimental avascular grafting procedures.This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (FNRS-PDR Convention T.0077.14, T,l,vie Grant no. 7.6515.16F awarded to DDM and Grant 5/4/150/5 awarded to M.M.D. [CAA is FRS-FNRS research associate]), Fonds Sp,ciaux de Recherche, and Fondation St Luc, Foundation Against Cancer, and donations from the Ferrero family. None of the authors have any competing interests to declare.