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Safety assessment of bone marrow derived MSC grown in platelet-rich plasma

机译：富含血小板的血浆中骨髓来源的MSC的安全性评估

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The injection of endothelial progenitor cells and mononuclear cells derived from bone marrow at the ischemic region of peripheral artery disease patients is reported to be effective for therapeutic angiogenesis; however, these cell therapies require large amounts of bone marrow to obtain sufficient numbers of cells. To solve this problem, we attempted to culture bone-marrow-derived mesenchymal stem cells (BM-MSC), which are supposed to secrete several cytokines that promote angiogenesis. We also focused on using platelet-rich plasma (PRP) as a supplement for cell culture instead of fetal bovine serum. Human BM-MSC obtained from healthy volunteers expanded rapidly when cultured with 10% PRP prepared from their own blood. FACS analysis revealed that these cultured human MSC were homogeneous populations, and chromosomal analysis showed a normal karyotype. Moreover, the angiogenetic effect was apparent two weeks after human BM-MSC were injected into the ischemic muscle in SCID mice. Tumor formation was not detected three months after injection into SCID mice either subcutaneously or intramuscularly. To simulate clinical settings, canine BM-MSC were grown with canine PRP and injected into their ischemic muscles. We confirmed that donor cells existed in situ two and six weeks after operation without any side effects. These results suggest that cultured human BM-MSC can be a promising cell source for therapeutic angiogenesis.
机译：据报道，在外周动脉疾病患者的缺血区域注射内皮祖细胞和骨髓来源的单核细胞可有效治疗血管生成。然而，这些细胞疗法需要大量的骨髓以获得足够数量的细胞。为了解决这个问题，我们尝试培养骨髓间充质干细胞（BM-MSC），该细胞应分泌几种促进血管生成的细胞因子。我们还专注于使用富含血小板的血浆（PRP）代替胎牛血清作为细胞培养的补充剂。从健康志愿者那里获得的人BM-MSC与自己血液中制备的10％PRP一起培养时迅速扩增。 FACS分析表明，这些培养的​​人MSC是同质种群，染色体分析显示出正常的核型。此外，在SCID小鼠的局部缺血肌肉中注射人BM-MSC两周后，血管生成作用明显。皮下或肌内注射入SCID小鼠三个月后未检测到肿瘤形成。为了模拟临床环境，将犬BM-MSC与犬PRP一起培养并注射到其缺血性肌肉中。我们确认手术后两周和六周原位存在供体细胞，没有任何副作用。这些结果表明，培养的人BM-MSC可以是治疗血管生成的有希望的细胞来源。

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