Direct contact with endoderm-like cells efficiently induces cardiac progenitors from mouse and human pluripotent stem cells.
Direct contact with endoderm-like cells efficiently induces cardiac progenitors from mouse and human pluripotent stem cells.
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DOI：
10.1371/journal.pone.0046413
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Kwon C
中科院分区：
文献类型：
--
作者：
Uosaki H;Andersen P;Shenje LT;Fernandez L;Christiansen SL;Kwon C
多能干细胞衍生的心脏祖细胞（CPCs）已成为体外研究心脏发生的有力工具和心脏再生医学的潜在细胞来源。然而，由于细胞系的变化，现有的诱导CPCs的方法效率不高，或者需要高成本的细胞因子进行大量优化。基于我们的体内观察，早期内胚层细胞与新生的心前中胚层保持接触，我们假设与内胚层的直接物理接触促进了多能细胞诱导CPCs。为了验证这一假设，我们将小鼠胚胎干细胞（ES）与内胚层细胞系End2通过共聚集或End2条件培养基共培养。共聚集导致Flk1+ PDGFRa+ CPCs以剂量依赖性的方式强烈诱导，但条件培养基没有，表明这一过程需要直接接触。为了确定与End2细胞的直接接触是否也促进了心脏祖细胞的诱导，我们使用了几种小鼠ES和诱导多能（iPS）细胞系，这些细胞系在CPC谱系标记物Nkx2.5或Isl1的调控下表达荧光蛋白。与早期的数据一致，与End2细胞共聚集可有效诱导Nkx2.5+和Isl1+心肌细胞，导致心肌细胞跳动。此外，与End2细胞的共聚集极大地促进了人类ES细胞诱导KDR+ PDGFRa+ CPCs。我们的共聚集方法为从小鼠和人多能性细胞中诱导CPCs提供了一种高效、简单和经济的方法。
Pluripotent stem cell–derived cardiac progenitor cells (CPCs) have emerged as a powerful tool to study cardiogenesis in vitro and a potential cell source for cardiac regenerative medicine. However, available methods to induce CPCs are not efficient or require high-cost cytokines with extensive optimization due to cell line variations. Based on our in-vivo observation that early endodermal cells maintain contact with nascent pre-cardiac mesoderm, we hypothesized that direct physical contact with endoderm promotes induction of CPCs from pluripotent cells. To test the hypothesis, we cocultured mouse embryonic stem (ES) cells with the endodermal cell line End2 by co-aggregation or End2-conditioned medium. Co-aggregation resulted in strong induction of Flk1+ PDGFRa+ CPCs in a dose-dependent manner, but the conditioned medium did not, indicating that direct contact is necessary for this process. To determine if direct contact with End2 cells also promotes the induction of committed cardiac progenitors, we utilized several mouse ES and induced pluripotent (iPS) cell lines expressing fluorescent proteins under regulation of the CPC lineage markers Nkx2.5 or Isl1. In agreement with earlier data, co-aggregation with End2 cells potently induces both Nkx2.5+ and Isl1+ CPCs, leading to a sheet of beating cardiomyocytes. Furthermore, co-aggregation with End2 cells greatly promotes the induction of KDR+ PDGFRa+ CPCs from human ES cells. Our co-aggregation method provides an efficient, simple and cost-effective way to induce CPCs from mouse and human pluripotent cells.
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影响因子：
4.8
作者：
Moretti, Alessandra;Bellin, Milena;Laugwitz, Karl-Ludwig
通讯作者：
Laugwitz, Karl-Ludwig
影响因子：
5.2
作者：
Arrell, D. Kent;Niederlaender, Nicolas J.;Terzic, Andre
通讯作者：
Terzic, Andre
DOI：
10.1073/pnas.0704044104
发表时间：
2007-06-26
影响因子：
11.1
作者：
Kwon, Chulan;Arnold, Joshua;Srivastava, Deepak
通讯作者：
Srivastava, Deepak
影响因子：
25
作者：
Madisen L;Zwingman TA;Sunkin SM;Oh SW;Zariwala HA;Gu H;Ng LL;Palmiter RD;Hawrylycz MJ;Jones AR;Lein ES;Zeng H
通讯作者：
Zeng H
影响因子：
64.5
作者：
Takahashi, Kazutoshi;Yamanaka, Shinya
通讯作者：
Yamanaka, Shinya