Determination of cell types and numbers during cardiac development in the neonatal and adult rat and mouse
Determination of cell types and numbers during cardiac development in the neonatal and adult rat and mouse
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DOI：
10.1152/ajpheart.00514.2007
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发表时间：
2007-09-01
影响因子：
4.8
通讯作者：
Baudino, Troy A.
中科院分区：
文献类型：
--
作者：
Banerjee, Indroneal;Fuseler, John W.;Baudino, Troy A.
心脏成纤维细胞、肌细胞、内皮细胞和血管平滑肌细胞是心脏的主要细胞成分。本研究的目的是观察成年动物在新生儿早期发育过程中心肌细胞群的变化，并观察不同种属（特别是大鼠和小鼠）心肌细胞群的任何变化。从新生儿和成年发育阶段的小鼠或大鼠中分离整个心脏，并通过连续胶原酶消化制备单细胞悬液。对异质细胞群进行特定细胞类型的免疫标记，并使用荧光激活细胞分选（FACS）进行分析。此外，分离、固定并切片左心室、右心室和间隔，用于形态测定分析。还使用FACS分析了这些相同的心脏区域。我们观察到成年小鼠心肌由大约56%的肌细胞、27%的成纤维细胞、7%的内皮细胞和10%的血管平滑肌细胞组成。此外，我们的形态测定和流式细胞仪的数据表明，在三个地区的检查相似的百分比。在小鼠新生儿心脏发育过程中，我们观察到心脏成纤维细胞的数量显着增加，并在新生儿发育后期（第15天）的肌细胞百分比下降。最后，在发育过程中大鼠心脏的流式细胞仪分析显示类似的结果，在发展过程中心脏成纤维细胞的增加，但是，在大鼠中的细胞群显着不同，从小鼠中观察到的。总之，这些数据使我们能够建立一个心肌的稳态模型，可以与遗传和心脏病模型进行比较。
Cardiac fibroblasts, myocytes, endothelial cells, and vascular smooth muscle cells are the major cellular constituents of the heart. The aim of this study was to observe alterations in myocardial cell populations during early neonatal development in the adult animal and to observe any variations of the cardiac cell populations in different species, specifically, the rat and mouse. Whole hearts were isolated from either mice or rats during the neonatal and adult stages of development, and single cell suspensions were prepared via sequential collagenase digestion. Heterogeneous cell populations were immuno-labeled for specific cell types and analyzed using fluorescence-activated cell sorting (FACS). In addition, the left ventricle, right ventricle, and septa were isolated, fixed, and sectioned for morphometric analyses. These same cardiac regions were also analyzed using FACS. We observed that the adult murine myocardium is composed of similar to 56% myocytes, 27% fibroblasts, 7% endothelial cells, and 10% vascular smooth muscle cells. Moreover, our morphometric and FACS data demonstrated similar percentages in the three regions examined. During murine neonatal cardiac development, we observed a marked increase in numbers of cardiac fibroblasts and a resultant decrease in percentages of myocytes in late neonatal development (day 15). Finally, FACS analyses of the rat heart during development displayed similar results in relation to increases in cardiac fibroblasts during development; however, cell populations in the rat differed markedly from those observed in the mouse. Taken together, these data enabled us to establish a homeostatic model for the myocardium that can be compared with genetic and cardiac disease models.