Department of Plastic and Reconstructive Surgery, Shanghai Key Laboratory of Tissue Engineering, Shanghai 9th People's Hospital, Shanghai Jiao Tong University, 639 Zhi Zao Ju Road, Shanghai, 200011, China.
Stem Cell Res Ther. 2018 Nov 8;9(1):294. doi: 10.1186/s13287-018-1014-y.
The proangiogenic capacity of adipose tissue and its derivatives has been demonstrated in a variety of studies. The paracrine mechanism of the cellular component is considered to play a critical role in the regenerative properties of these tissues. However, cell-based therapy for clinical application has been hindered by limitations such as safety, immunogenicity issues, and difficulties in cell preservation, transportation, and phenotype control. In the current study, we aimed to produce a cell-free extract directly from human fat tissue and evaluate its potential therapeutic efficacy.
We developed a novel physical approach to produce a cell-free aqueous extract from human fat tissue (fat extract (FE)). The therapeutic potential of FE was investigated in the ischemic hindlimb model of nude mice. After establishment of hindlimb ischemia with ligation of the left femoral artery and intramuscular injection of FE, blood perfusion was monitored at days 0, 7, 14, 21, and 28. Tissue necrosis and capillary density were evaluated. Enzyme-linked immunosorbent assay was used to analyze the growth factors contained in FE. Moreover, the proliferation, migration, and tube formation ability were tested on human umbilical vein endothelial cells (HUVECs) in vitro when treated with FE. The proangiogenic ability of FE was further assessed in an in-vivo Matrigel plug assay.
FE was prepared and characterized. The intramuscular injection of FE into the ischemic hindlimb of mice attenuated severe limb loss and increased blood flow and capillary density of the ischemic tissue. Enzyme-linked immunosorbent assay showed that FE contained high levels of various growth factors. When added as a cell culture supplement, FE promoted HUVEC proliferation, migration, and tube formation ability in a dose-dependent manner. The subcutaneous injection of Matrigel infused with FE enhanced vascular formation.
We developed a novel cell-free therapeutic agent, FE, produced from human adipose tissue. FE was able to attenuate ischemic injury and stimulate angiogenesis in ischemic tissues. This study indicates that FE may represent a novel cell-free therapeutic agent in the treatment of ischemic disorders.
脂肪组织及其衍生物的促血管生成能力已在多种研究中得到证实。细胞成分的旁分泌机制被认为在这些组织的再生特性中起着关键作用。然而，细胞治疗因其安全性、免疫原性问题以及细胞保存、运输和表型控制困难等限制，难以应用于临床。在本研究中，我们旨在直接从人体脂肪组织中产生无细胞提取物，并评估其潜在的治疗效果。
我们开发了一种从人体脂肪组织中产生无细胞水提物的新型物理方法（脂肪提取物（FE））。FE 的治疗潜力在裸鼠缺血后肢模型中进行了研究。通过结扎左股动脉和肌肉内注射 FE 建立后肢缺血后，在第 0、7、14、21 和 28 天监测血液灌注。评估组织坏死和毛细血管密度。采用酶联免疫吸附试验分析 FE 中所含的生长因子。此外，当用 FE 处理时，在体外测试人脐静脉内皮细胞（HUVEC）的增殖、迁移和管形成能力。FE 的促血管生成能力进一步在体内 Matrigel plugs 实验中进行评估。
FE 已制备并进行了表征。FE 肌肉内注射到小鼠缺血后肢可减轻严重的肢体丧失并增加缺血组织的血流和毛细血管密度。酶联免疫吸附试验显示 FE 含有高水平的各种生长因子。当作为细胞培养补充剂添加时，FE 以剂量依赖性方式促进 HUVEC 的增殖、迁移和管形成能力。FE 皮下注射到 Matrigel 中可增强血管形成。
我们开发了一种从人体脂肪组织中产生的新型无细胞治疗剂 FE。FE 能够减轻缺血损伤并刺激缺血组织中的血管生成。这项研究表明，FE 可能代表治疗缺血性疾病的新型无细胞治疗剂。
