Optimization of EDU Pulse Method for Tracing the Migration of Mouse Second Heart Field Cells

doi:10.13560/j.cnki.biotech.bull.1985.2018-0518

Abstract

Abstract: Outflow tract（OFT）malformation accounts for about 30% of detected human congenital heart diseases at birth. OFT and right ventricle are developed from second heart field（SHF）. Deployment and migration of SHF cells to heart tube is essential for sufficient lengthening of heart tube and subsequent heart morphogenesis. However,the development,migration and deployment process of SHF is still unclear. Establishing a method of marking and tracing the migration of second heart field cells in the mouse heart is of great significance in elucidating the cellular biological processes of SHF migration and deployment in mammalian hearts. Firstly,we adapted EDU（5-ethynyl-2'-deoxyuridine）pulse-chase assay with dose（10 mg/kg）reported in the literature,and the most embryo were not labeled by EDU. Following,we optimized the dosage and pulse time by analyzing the labeling effects of EDU in SHF cells and the migration of EDU-labeled SHF cells to OFT. The dosage of 40 mg/kg demonstrated effective labeling on SHF cells（31 embryos from 6 pregnant mice were efficiently labeled）,and no toxicity to embryos and pregnant mice. Moreover,2 h- and 4 h- EDU pulse period showed no difference in EDU labeling efficiency and the migratory distance to the distal OFT. Taking above results together,the 40 mg/kg EDU pulse dose and 2 h EDU pulse period presented effective EDU labeling of SHF cells in embryos,and many EDU labeling SHF cells can be detected in the distal OFT after 19 h tracing period.

Key words: second heart field cells, EDU, cell migration, mouse

YUAN Bai-yin, LIU Zhong-ying. Optimization of EDU Pulse Method for Tracing the Migration of Mouse Second Heart Field Cells[J]. Biotechnology Bulletin, 2018, 34(12): 84-89.

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