Derivation and characterization of human fetal MSCs: An alternative cell source for large-scale production of cardioprotective microparticles
Derivation and characterization of human fetal MSCs: An alternative cell source for large-scale production of cardioprotective microparticles
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DOI：
10.1016/j.yjmcc.2009.12.021
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发表时间：
2010-06-01
影响因子：
5
通讯作者：
Lim, Sai Kiang
中科院分区：
文献类型：
--
作者：
Lai, Ruenn Chai;Arslan, Fatih;Lim, Sai Kiang
间充质干细胞（MSC）移植的治疗效果越来越被认为是由MSC分泌介导的。我们以前已经证明，人胚胎干细胞衍生的骨髓间充质干细胞（hESC-MSCs）产生心肌保护微粒在猪模型的心肌缺血/再灌注（MI/R）损伤。由于临床级人ESC的安全性和可用性仍然是一个问题，因此评估了来自胎儿组织来源的MSC作为替代品。在这里，我们从三个前三个月流产胎儿的肢体，肾脏和肝脏组织中获得了五种MSC培养物，就像我们之前描述的hESC衍生的MSC一样;它们具有高度可扩展性，并且具有相似的端粒酶活性。对于MI/R损伤的猪模型，每个细胞系具有产生至少10（16-19）个细胞或10（7-10）个剂量的心脏保护分泌物的潜力。与先前描述的胎儿MSC不同，它们不表达多能性相关标记物，如Oct 4。Nanog或Tra 1 -60。它们表现出典型的MSC表面抗原谱，并在体外分化为脂肪细胞、骨细胞和软骨细胞。通过微阵列和qRT-PCR的全局基因表达分析揭示了典型的MSC基因表达谱，其在五种胎儿MSC培养物中与hESC-MSC的基因表达谱高度相关（r（2）> 0.90）。与hESC-MSC一样，它们产生的分泌物在MI/R损伤的小鼠模型中具有心脏保护作用。分泌物的HPLC分析揭示了流体动力学半径为50-65 nm的微粒群的存在。这种纯化的微粒群在与粗分泌物的1/10剂量相似的情况下具有心脏保护作用。(C)2010爱思唯尔有限公司版权所有。
The therapeutic effects of mesenchymal stem cells (MSCs) transplantation are increasingly thought to be mediated by MSC secretion. We have previously demonstrated that human ESC-derived MSCs (hESC-MSCs) produce cardioprotective microparticles in pig model of myocardial ischemia/reperfusion (MI/R) injury. As the safety and availability of clinical grade human ESCs remain a concern, MSCs from fetal tissue sources were evaluated as alternatives. Here we derived five MSC cultures from limb, kidney and liver tissues of three first trimester aborted fetuses and like our previously described hESC-derived MSCs; they were highly expandable and had similar telomerase activities. Each line has the potential to generate at least 10(16-19) cells or 10(7-10) doses of cardioprotective secretion for a pig model of MI/R injury. Unlike previously described fetal MSCs, they did not express pluripotency-associated markers such as Oct4. Nanog or Tra1-60. They displayed a typical MSC surface antigen profile and differentiated into adipocytes, osteocytes and chondrocytes in vitro. Global gene expression analysis by microarray and qRT-PCR revealed a typical MSC gene expression profile that was highly correlated among the five fetal MSC cultures and with that of hESC-MSCs (r(2) > 0.90). Like hESC-MSCs, they produced secretion that was cardioprotective in a mouse model of MI/R injury. HPLC analysis of the secretion revealed the presence of a population of microparticles with a hydrodynamic radius of 50-65 nm. This purified population of microparticles was cardioprotective at similar to 1/10 dosage of the crude secretion. (C) 2010 Elsevier Ltd. All rights reserved.