Concerted Regulation of CD34 and CD105 Accompanies Mesenchymal Stromal Cell Derivation from Human Adventitial Stromal Cell
Concerted Regulation of CD34 and CD105 Accompanies Mesenchymal Stromal Cell Derivation from Human Adventitial Stromal Cell
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DOI：
10.1089/scd.2012.0263
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发表时间：
2013-03-01
影响因子：
4
通讯作者：
Dong, Jun
中科院分区：
文献类型：
--
作者：
Braun, Julian;Kurtz, Andreas;Dong, Jun
间充质基质细胞(MSC)在创新的治疗应用方面得到了广泛的研究。MSC在体外具有贴壁增殖、免疫表型特异性和多能性的特点，而体内MSC的前体细胞被描述为血管周围细胞。MSC祖细胞是否在体外培养中获得MSC特性尚不清楚。这个问题可以通过分析在早期体外培养阶段，即MSC衍生阶段发生的细胞特性变化来实验地解决。在这里，我们检测了在脂肪组织来源的MSC(AT-MSC)的MSC衍生阶段，用于MSC表征的表面标志物(如CD34、CD105、CD146和CD271)在形态、增殖和表达方面的动态变化。用多参数流式细胞术鉴定了CD34(+)CD146(-)CD271(+/-)外膜基质细胞样细胞(ADSC)、CD34(-)CD146(+)CD271(+/-)周细胞样细胞(PC)和CD34(+)CD31(+)CD146(+)内皮细胞。在MSC培养条件下，只有ADSC而不是PC能分化为MSC。培养第4天，ADSC呈成纤维样细胞，CD105、CD146、CD271表达上调。在这种表型转变之后，ADSC开始增殖并下调CD34的表达。在我们的研究中，我们证明ADSC是比PC更多的克隆性AT-MSC前体细胞。此外，我们首次使用多参数流式细胞术分析了在MSC衍生阶段从MSC前体细胞到体外MSC的表型转变。因此，我们提出了一个模型，描述了ADSC在体外培养中如何从头获得典型的MSC形态，并伴随着对表面标志表达的协同调节。
Mesenchymal stromal cells (MSC) have been intensively studied for innovative therapeutic applications. MSC in vitro are characterized by plastic-adherent proliferation, their specific immunophenotype and multipotency, whereas MSC progenitors in vivo are described as perivascular cells. Whether MSC progenitors acquire in vitro MSC characteristics upon in vitro culture is still unclear. This question can be experimentally accessed by analyzing changes in cellular properties that occur during the early in vitro culture phase, the MSC derivation phase. Here, we examined dynamics in morphology, proliferation, and expression of surface markers used for MSC characterization (such as CD34, CD105, CD146, and CD271) in tight kinetics during the MSC derivation phase of adipose tissue-derived MSC (AT-MSC). Using multiparametric flow cytometry, we identified 3 major ex vivo stromal vascular cell subsets: CD34(+)CD146(-)CD271(+/-) adventitial stromal cell-like cells (AdSC), CD34(-)CD146(+)CD271(+/-) pericyte-like cells (PC), and CD34(+)CD31(+)CD146(+) endothelial cells. Of these subsets, only AdSC, but not PC gave rise to MSC under MSC culture conditions. At day 4 of culture, AdSC became fibroblastoid and upregulated CD105, CD146, and CD271. Following this phenotypic transition, AdSC commenced proliferation and downregulated CD34. In our study, we demonstrate that AdSC are more clonogenic AT-MSC progenitors than PC. Moreover, we, for the first time have dissected the phenotypic transitions from MSC progenitors to in vitro MSC during the MSC derivation phase using multiparametric flow cytometry. Hence, we propose a model describing how de novo acquisition of the typical MSC morphology by AdSC is accompanied by concerted regulation of surface marker expression upon in vitro culture.