Abstract
Cell sorting via flow cytometry is a powerful tool to select subpopulations of cells in many biological fields. Selection of fertilisation-prone sperm is a critical step to ensure a stable and high fertilisation rate in in vitro fertilisation (IVF). However, a combination of conventional cell sorting and IVF system has not been established because of severe mechanical damages to the sperm during the sorting process. A cell sorter with microfluidics chip technology that lessens cell damage during cell sorting may address this problem. We evaluated the effects of microfluidics chip cell sorting on the sperm using the parameters, such as motility and fertility, and found this cell sorting method had minimal harmful effect on the sperm. Then, sperm were selected by a marker for acrosome reaction and showed higher fertilisation rate than that of the population of acrosome-intact sperm. Embryo derived from these sperm developed normally. These results indicated that microfluidics chip cell sorting can select fertile sperm to improve IVF technique.
Highlights
Cell sorting via flow cytometry is a powerful tool to select subpopulations of cells in many biological fields
Sperm were sorted by the microfluidics chip cell sorter based on the signal distribution of forward scattered light (FSC) and side scattered light (SSC) (Fig. 1A)
Sperm selection by flow cytometry is usually used for measuring DNA content[11,12,13]
Summary
Cell sorting via flow cytometry is a powerful tool to select subpopulations of cells in many biological fields. We evaluated the effects of microfluidics chip cell sorting on the sperm using the parameters, such as motility and fertility, and found this cell sorting method had minimal harmful effect on the sperm. Sperm were selected by a marker for acrosome reaction and showed higher fertilisation rate than that of the population of acrosome-intact sperm Embryo derived from these sperm developed normally. These results indicated that microfluidics chip cell sorting can select fertile sperm to improve IVF technique. Cell sorting using flow cytometry is a powerful method for selecting subpopulation of cells in many biological studies involving various indicators of fluorescence proteins, antibodies and compounds. We first examined whether the microfluidics chip cell sorter is useful for sperm selection by using the parameters such as motility, fertility and developmental ability of the collected sperm. We selected acrosome-reacted sperm using fluorescein isothiocyanate (FITC, a fluorescence compound)-labelled peanut agglutinin (PNA) to demonstrate the efficacy of this system
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