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细胞遗传标记可用于鉴定和追踪小鼠中的转基因血小板。

Genetic Labeling of Cells Allows Identification and Tracking of Transgenic Platelets in Mice.

机构信息

Department of Vascular and Endovascular Surgery, Experimental Vascular Medicine, Heinrich-Heine University Medical Center, 40225 Düsseldorf, Germany.

出版信息

Int J Mol Sci. 2021 Apr 2;22(7):3710. doi: 10.3390/ijms22073710.

Abstract

BACKGROUND

The use of knock-out mouse models is crucial to understand platelet activation and aggregation.

METHODS

Analysis of the global double fluorescent Cre reporter mouse that has been crossbred with the megakaryocyte/platelet specific mouse.

RESULTS

Platelets show bright ( negative) and ( positive) fluorescence. However, a small proportion of leukocytes was positive for fluorescence in positive mice. In mice, platelets, and megakaryocytes can be tracked by their specific fluorescence in blood smear, hematopoietic organs and upon thrombus formation. No differences in platelet activation and thrombus formation was observed between positive and negative mice. Furthermore, hemostasis and in vivo thrombus formation was comparable between genotypes as analyzed by intravital microscopy. Transplantation studies revealed that bone marrow of mice can be transferred to mice.

CONCLUSIONS

The reporter mouse is an appropriate model for real-time visualization of platelets, the analysis of cell morphology and the identification of non-recombined platelets. Thus, mice are important for the analysis of platelet-specific knockout mice. However, a small proportion of leukocytes exhibit fluorescence. Therefore, the analysis of platelets beyond hemostasis and thrombosis should be critically evaluated when recombination of immune cells is increased.

摘要

背景

敲除小鼠模型的使用对于理解血小板的激活和聚集至关重要。

方法

分析已与巨核细胞/血小板特异性小鼠杂交的全局双荧光 Cre 报告小鼠。

结果

血小板显示出明亮的(阴性)和(阳性)荧光。然而,在阳性小鼠中,一小部分白细胞对荧光呈阳性。在 小鼠中,血小板和巨核细胞可以通过其在血涂片、造血器官和血栓形成时的特异性荧光来追踪。在阳性和阴性小鼠之间未观察到血小板激活和血栓形成的差异。此外,通过活体显微镜分析,止血和体内血栓形成在基因型之间具有可比性。移植研究表明, 小鼠的骨髓可以转移到 小鼠中。

结论

该 报告小鼠是实时可视化血小板、分析细胞形态和识别未重组血小板的合适模型。因此, 小鼠对于血小板特异性敲除小鼠的分析很重要。然而,一小部分白细胞显示出 荧光。因此,当免疫细胞的重组增加时,应仔细评估超越止血和血栓形成的血小板分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f3f3/8037568/c24f2c77bbbb/ijms-22-03710-g001.jpg

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