Impact of Different Method in Sperm Membrane Treatment on Exogenous Plasmid Transgenic Efficiency by Sperm

doi:10.3969/j.issn.1674-5817.2013.04.002

Abstract

Abstract: Objective To explore the effect of three methods in removing mouse sperm membrane including ultrasound, NaOH and lysolecithin on the efficiency of transgenic by intracytoplasmic sperm injection (ICSI). Methods The mouse sperms were treated with the three methods above mentioned. Both of sperm membrane and DNA integrity of the sperm cells were evaluated by flow cytometry after died with Transgreen and propidium iodide (PI). The treated sperms were incubated with GFP-carrying plasmid and then conducted ICSI. The effect of these methods on efficiency of transgenic was evaluated by the results of fertilization cleavage and blastocyst with fluorescence. Results The rate of removing sperm membrane by lysolecithin method was higher than that of ultrasound and NaOH methods (99.2% vs. 94.7%, 90.9%)(P<0.05), but the rates of sperm DNA damage, the development of embryos from ICSI and blastocyst with fluorescence were similar among these methods (P>0.05). Conclusion The three methods of mouse membrane treatment could be used to improve mouse transgenic efficiency by ICSI.

Key words: Method, Sperm membrane treatment, ICSI, DNA integrity, Transgenic efficiency, Mouse

CLC Number:

Q95-33

JIA Xiao-feng, XU Yan, LIU Miao, WANG Liang, SHI Hui-juan. Impact of Different Method in Sperm Membrane Treatment on Exogenous Plasmid Transgenic Efficiency by Sperm[J]. Laboratory Animal and Comparative Medicine, 2013, 33(4): 256-260.

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