Human Mesenchymal Stromal Cells: Identifying Assays to Predict Potency for Therapeutic Selection
Human Mesenchymal Stromal Cells: Identifying Assays to Predict Potency for Therapeutic Selection
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DOI：
10.5966/sctm.2012-0099
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发表时间：
2013-02-01
影响因子：
6
通讯作者：
Young, Pampee P.
中科院分区：
文献类型：
--
作者：
Deskins, Desirae L.;Bastakoty, Dikshya;Young, Pampee P.
多能间充质基质细胞（MSC）具有修复和再生受损组织的潜力，使其成为基于细胞的治疗的有吸引力的候选者。为了最大限度地提高MSC的疗效，必须预测其治疗能力，以便只使用最好的细胞。我们的目标是确定可行且可重复的体外试验来预测MSC的效价。我们从10个正常人骨髓样本中产生细胞系，并使用国际细胞治疗学会的最低标准将其定义为MSC：塑性粘附，适当的表面标记物表达和三系分化。每种MSC系进一步通过其生长、增殖和活力来表征，如分别通过细胞计数、溴脱氧尿苷掺入和细胞ATP水平所确定的。为了确定这些测试是否可靠地预测MSC的治疗倾向，将几种细胞系植入体内，以检查它们在使用聚乙烯醇海绵的良好建立的小鼠伤口模型中移植和形成肉芽组织的能力。MSC在海绵中的长期植入通过海绵切片中人类特异性Alu基因的存在来量化。还对切片的增殖细胞、血管分布和肉芽组织形成进行染色，以确定成功的植入和修复。我们发现，在体外测试的组合中的高性能准确地预测了哪些线在体内功能良好。这些研究结果表明，可靠的和可重复的体外测定可用于测量MSC的治疗用途的功能潜力。干细胞翻译医学2013;2：151-158
Multipotent mesenchymal stromal cells (MSCs) have the potential to repair and regenerate damaged tissues, making them attractive candidates for cell-based therapies. To maximize efficacy of MSCs, prediction of their therapeutic abilities must be made so that only the best cells will be used. Our goal was to identify feasible and reproducible in vitro assays to predict MSC potency. We generated cell lines from 10 normal human bone marrow samples and used the International Society for Cellular Therapy's minimal criteria to define them as MSCs: plastic adherence, appropriate surface marker expression, and trilineage differentiation. Each MSC line was further characterized by its growth, proliferation, and viability as determined by cell count, bromodeoxyuridine incorporation, and cellular ATP levels, respectively. To determine whether these tests reliably predict the therapeutic aptitude of the MSCs, several lines were implanted in vivo to examine their capacity to engraft and form granulation tissue in a well-established murine wound model using polyvinyl alcohol sponges. Long-term engraftment of MSCs in the sponges was quantified through the presence of the human-specific Alu gene in sponge sections. Sections were also stained for proliferating cells, vascularity, and granulation tissue formation to determine successful engraftment and repair. We found that high performance in a combination of the in vitro tests accurately predicted which lines functioned well in vivo. These findings suggest that reliable and reproducible in vitro assays may be used to measure the functional potential of MSCs for therapeutic use. STEM CELLS TRANSLATIONAL MEDICINE 2013;2:151-158