Maijub John G, Boyd Nolan L, Dale Jacob R, Hoying James B, Morris Marvin E, Williams Stuart K
Cardiovascular Innovation Institute, Department of Surgery, University of Louisville School of Medicine, Louisville, KY, USA.
Cell Transplant. 2015;24(10):2029-39. doi: 10.3727/096368914X685401. Epub 2014 Nov 13.
Adipose-derived stromal vascular fraction (SVF) cells have been shown to self-associate to form vascular structures under both in vitro and in vivo conditions. The angiogenic (new vessels from existing vessels) and vasculogenic (new vessels through self-assembly) potential of the SVF cell population may provide a cell source for directly treating (i.e., point of care without further cell isolation) ischemic tissues. However the correct dosage of adipose SVF cells required to achieve a functional vasculature has not been established. Accordingly, in vitro and in vivo dose response assays were performed evaluating the SVF cell vasculogenic potential. Serial dilutions of freshly isolated rat adipose SVF cells were plated on growth factor reduced Matrigel and vasculogenesis, assessed as cellular tube-like network assembly, was quantified after 3 days of culture. This in vitro vasculogenesis assay indicated that rat SVF cells reached maximum network length at a concentration of 2.5 × 10(5) cells/ml and network maintained at the higher concentrations tested. The same concentrations of rat and human SVF cells were used to evaluate vasculogenesis in vivo. SVF cells were incorporated into collagen gels and subcutaneously implanted into Rag1 immunodeficient mice. The 3D confocal images of harvested constructs were evaluated to quantify dose dependency of SVF cell vasculogenesis potential. Rat- and human-derived SVF cells yielded a maximum vasculogenic potential at 1 × 10(6) and 4 × 10(6) cells/ml, respectively. No adverse reactions (e.g., toxicity, necrosis, tumor formation) were observed at any concentration tested. In conclusion, the vasculogenic potential of adipose-derived SVF cell populations is dose dependent.
脂肪来源的基质血管成分（SVF）细胞已被证明在体外和体内条件下均能自我缔合形成血管结构。SVF细胞群体的血管生成（从现有血管生成新血管）和血管发生（通过自我组装生成新血管）潜力可能为直接治疗（即无需进一步细胞分离的即时护理）缺血组织提供细胞来源。然而，实现功能性脉管系统所需的脂肪SVF细胞的正确剂量尚未确定。因此，进行了体外和体内剂量反应试验，以评估SVF细胞的血管发生潜力。将新鲜分离的大鼠脂肪SVF细胞进行系列稀释后接种在生长因子减少的基质胶上，培养3天后，对作为细胞管状网络组装评估的血管发生进行定量。这种体外血管发生试验表明，大鼠SVF细胞在浓度为2.5×10⁵个细胞/ml时达到最大网络长度，且在更高测试浓度下网络保持稳定。使用相同浓度的大鼠和人类SVF细胞评估体内血管发生。将SVF细胞掺入胶原凝胶中，然后皮下植入Rag1免疫缺陷小鼠体内。对收获的构建体进行三维共聚焦成像评估，以量化SVF细胞血管发生潜力的剂量依赖性。大鼠和人类来源的SVF细胞分别在1×10⁶和4×10⁶个细胞/ml时产生最大血管发生潜力。在任何测试浓度下均未观察到不良反应（如毒性、坏死、肿瘤形成）。总之,脂肪来源的SVF细胞群体的血管发生潜力是剂量依赖性的。
