Lineage tracing of cardiac explant derived cells.
Lineage tracing of cardiac explant derived cells.
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DOI：
10.1371/journal.pone.0001929
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发表时间：
2008-04-16
期刊：
影响因子：
3.7
通讯作者：
Galiñanes M
中科院分区：
文献类型：
--
作者：
Shenje LT;Field LJ;Pritchard CA;Guerin CJ;Rubart M;Soonpaa MH;Ang KL;Galiñanes M
培养的心脏外植体产生异质细胞群，包括独特的嗜酸性细胞群，在此描述为小圆心脏外植体衍生细胞（EDC）。本研究旨在探讨内分泌干扰物的来源、形态及致心功能。转基因MLC 2 v-Cre/ZEG和actin-eGFP小鼠用于体外和体内EDC的谱系追踪。C57 B16小鼠用作来自转基因心脏的EDC的细胞移植受体，以及用于EDC的一般表征。心脏特异性标志物的激活通过：免疫组织化学与明视野和免疫荧光显微镜，电子显微镜，PCR和RT-PCR分析。用钙瞬变研究进一步研究移植细胞的功能性植入。内分泌干扰物的产生高度依赖于培养外植体中血液来源的细胞或因子的保留。这些细胞在体外具有心肌细胞的一些特征，并在体内植入成人心脏中存活。然而，内分泌干扰物未能分化成功能性心肌细胞在体内的刺激诱发的细胞内钙瞬变的情况下移植到梗死周围区。本研究强调，仅基于一个参数（如形态学或免疫荧光）的阳性鉴定不足以鉴定成人心脏外植体衍生细胞的来源、命运和功能。
Cultured cardiac explants produce a heterogeneous population of cells including a distinctive population of refractile cells described here as small round cardiac explant derived cells (EDCs). The aim of this study was to explore the source, morphology and cardiogenic potential of EDCs. Transgenic MLC2v-Cre/ZEG, and actin-eGFP mice were used for lineage-tracing of EDCs in vitro and in vivo. C57B16 mice were used as cell transplant recipients of EDCs from transgenic hearts, as well as for the general characterisation of EDCs. The activation of cardiac-specific markers were analysed by: immunohistochemistry with bright field and immunofluorescent microscopy, electron microscopy, PCR and RT-PCR. Functional engraftment of transplanted cells was further investigated with calcium transient studies. Production of EDCs was highly dependent on the retention of blood-derived cells or factors in the cultured explants. These cells shared some characteristics of cardiac myocytes in vitro and survived engraftment in the adult heart in vivo. However, EDCs failed to differentiate into functional cardiac myocytes in vivo as demonstrated by the absence of stimulation-evoked intracellular calcium transients following transplantation into the peri-infarct zone. This study highlights that positive identification based upon one parameter alone such as morphology or immunofluorescene is not adequate to identify the source, fate and function of adult cardiac explant derived cells.
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