High passage number of stem cells adversely affects stem cell activation and myocardial protection
High passage number of stem cells adversely affects stem cell activation and myocardial protection
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DOI：
10.1097/01.shk.0000235087.45798.93
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发表时间：
2006-12-01
期刊：
影响因子：
3.1
通讯作者：
Meldrum, Daniel R.
中科院分区：
文献类型：
--
作者：
Crisostomo, Paul R.;Wang, Meijing;Meldrum, Daniel R.
祖细胞可塑性增强受损组织的正性重塑。我们和其他人以前已经表明，祖细胞可能限制凋亡和调节炎症的部分生产的生长因子。然而，最近的研究表明，祖细胞随着培养和传代时间的增加而衰老并失去其分化潜能。我们假设，小鼠骨髓间充质干细胞（MSC）对缺血/再灌注损伤的心脏保护在离体灌注大鼠心脏，和传代次数对MSC的激活和心脏保护有不利影响。分离成年雄性和雌性Sprague-Dawley大鼠心脏，通过Langendorff模型灌注，并进行缺血/再灌注。收获小鼠MSC，培养，悬浮于灌注液中，并在全局指数缺血前输注。将心脏分配为对照组或输注第3、5或10代MSC。此外，培养中的MSC通过缺氧和增加剂量的内毒素（脂多糖）来应激。间充质干细胞活化通过用酶联免疫吸附测定法测量血管内皮生长因子的产生来确定。所有数据均报告为平均值+/- SEM，并用双因素方差分析进行分析。如果P< 0.05，则认为差异显著。缺血前输注第3代小鼠MSC可减轻左心室发展压的降低，减弱舒张末压的升高，减轻+dP/dT和-dP/dT的降低。而第5代和第10代骨髓间充质干细胞对心肌的保护作用消失。虽然缺氧和脂多糖导致MSC的显著活化，但第3代MSC表现出比第5代和第10代MSC显著更大的血管内皮生长因子释放。急性鼠MSC输注在离体大鼠心脏中赋予保护作用。然而，高传代次数对MSC活化和保护具有不利影响。这预示着在可能的治疗用途之前，离体扩张有限。
Progenitor cell plasticity enhances positive remodeling of damaged tissue. We and others have previously shown that progenitor cells may limit apoptosis and modulate inflammation in part by the production of growth factors. However, recent studies suggest that progenitor cells senesce and lose their differentiation potential with increasing time in culture and passage. We hypothesize that murine bone marrow mesenchymal stem cells (MSCs) are cardioprotective against ischemia/ reperfusion injury in the isolated perfused rat heart, and that passage number has an adverse effect on MSC activation and cardioprotection. Adult male and female Sprague-Dawley rat hearts were isolated, perfused via Langendorff model, and subjected to ischemia/reperfusion. Mouse MSCs were harvested, cultured, suspended in perfusate, and infused before global index ischemia. Hearts were assigned to controls or infusion with passage 3, 5, or 10 MSCs. In addition, MSCs in culture were stressed by hypoxia and increasing doses of endotoxin (lipopolysaccharide). Mesenchymal stem cell activation was determined by measuring vascular endothelial growth factor production with enzyme-linked immunosorbent assay. All data are reported as mean +/- SEM and were analyzed with 2-way analysis of variance. Differences are considered significant if P< 0.05. Passage 3 murine MSC infusion in hearts before ischemia reduced the depression of left ventricular developed pressure, attenuated the increase of end-diastolic pressure, and reduced the depression of +dP/dT and -dP/dT. However, the MSC protective effect disappeared in hearts infused with passage 5 and passage 10 MSCs. Although hypoxia and lipopolysaccharide resulted in significant activation of MSCs, passage 3 MSCs demonstrated significantly greater vascular endothelial growth factor release than passage 5 and 10 MSCs. Acute murine MSC infusion confers protection in isolated rat hearts. However, high passage number has an adverse effect on MSC activation and protection. This portends limited ex vivo expansion before possible therapeutic use.