Establishment of optimal tissue regenerating procedures using monocyte-derived multipotential cells for tissue regeneration

我们最近发现了一种原始的人类细胞群，称为单核细胞衍生多潜能细胞(MOMC)，它具有成纤维细胞样的形态，CD14、CD45、CD34和I型胶原呈独特的阳性表型。MOMC含有能够分化为多种非吞噬细胞的前体细胞，包括骨、软骨、脂肪、骨骼和心肌、神经元和内皮细胞。本研究的目的是评价MOMC移植在组织再生中的作用，并建立适合组织再生治疗的MOMC的最佳培养条件。1)动物模型中MOMC移植的效果评价采用两种不同的大鼠模型，包括脑梗塞和皮瓣移植。大鼠左侧大脑中动脉闭塞1h后，于第7天行大鼠大脑中动脉微血管内移植。…移植组大鼠神经功能改善巨噬细胞移植组和单纯培养液移植组大鼠的MOMC数明显增加，而对照组大鼠无明显变化。免疫组织化学分析显示，与对照组相比，移植MOMC的大鼠新生血管数量显著增加。移植的MOMC被证实整合到血管壁中，并分化为内皮细胞。从大鼠皮瓣模型中获得了关于MOMC功能改善和体内向内皮细胞分化的基本一致的发现。2)建立了生成人MOMC的最佳培养条件。我们先前发现，在激活的血小板来源的可溶性因子(S)的存在下，在纤维连接蛋白包裹的塑料平板上，MOMC可以在人外周血单个核细胞的培养中浓缩。通过在存在或不存在分子量为30,000的候选分子的情况下培养循环单核细胞来筛选具有这种能力的潜在的血小板衍生因子，这些分子在血小板激活时被释放。因此，SDF-1被确定为促进MOMC生成的一个因素。在此基础上，我们成功地开发了一种体外方法，通过在含有外源性SDF-1的重组纤维连接蛋白上培养循环单核细胞来产生大量的人MOMC。这些发现共同表明，MOMC移植是一种有前景的再生受损组织的策略。较少

We recently discovered a primitive human cell population called monocyte-derived multipotential cells (MOMC), which has a fibroblast-like morphology and a unique phenotype positive for CD14, CD45, CD34, and type I collagen. MOMC contain progenitors with capacity to differentiate into a variety of non-phagocytes, including bone, cartilage, fat, skeletal and cardiac muscle, neuron, and endothelium. This study was aimed to assess efficacy of MOMC transplantation in tissue regeneration and to establish optimal culture conditions that generate MOMC for tissue regeneration therapy.1)Evaluation of efficacy of MOMC transplantation in animal modelsEfficacy of transplantation of syngeneic MOMC was assessed using 2 different rat models, including cerebral infarction and skin flap procedure. Rats were subjected to 1 hour of left middle cerebral artery occlusion and received intra-cranial transplantation of MOMCs at day 7. Neurological function assessed by Corner test was improved in rats transplan … More ted of MOMC, but not in control rats transplanted of macrophages or medium alone. In immunohistochemical analysis, the number of newly formed blood vessels was significantly increased in MOMC-transplanted rats, compared with controls rats. Transplanted MOMC were confirmed to be incorporated in blood vessel wall and differentiate into endothelial cells. A principally concordant finding regarding functional improvement and in vivo differentiation of MOMC to endothelial cells was obtained from rat skin flap model.2)Establishment of optimal culture conditions that generate human MOMCWe previously found that MOMC can be enriched in culture of human peripheral blood mononuclear cells on fibronectin-coated plastic plates in the presence of soluble factor(s) derived from activated platelets. Potential platelet-derived factors with such capacity were screened by culturing circulating monocytes in the presence or absence of candidate molecules with molecular weight < 30,000, which were released upon platelet activation. As a result, SDF-1 was identified as a factor that promotes MOMC generation. Based on this, we have successfully developed an ex vivo procedure that generates a large number of human MOMC by culturing circulating monocytes on recombinant fibronectin with exogenous SDF-1.These findings together indicate that MOMC transplantation is a promising strategy that regenerates the damaged tissue. Less