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. 2023 Jul 17;26(8):107356.
doi: 10.1016/j.isci.2023.107356. eCollection 2023 Aug 18.

Design of a gradient-rheotaxis microfluidic chip for sorting of high-quality Sperm with progressive motility

Chung-Hsien Huang  1   2 , Ching-Hung Chen  3 , Teng-Kuan Huang  1   2 , Farn Lu  3 , Jack Yu Jen Huang  3   4 , Bor-Ran Li  1   2   5   6
Affiliations

Design of a gradient-rheotaxis microfluidic chip for sorting of high-quality Sperm with progressive motility

Chung-Hsien Huang et al. iScience. .

Abstract

Assisted reproductive technology (ART) is an important invention for the treatment of human infertility, and the isolation of high-quality sperm with progressive motility is one of the most critical steps that eventually affect the fertilization rate. Conventional sperm separation approaches include the swim-up method and density gradient centrifugation. However, the quality of isolated sperm obtained from both approaches can still be improved by improving sorted sperm motility, minimizing the DNA fragmentation rate, and removing abnormal phenotypes. Here, we report a Progressive Sperm Sorting Chip (PSSC) for high-quality sperm isolation. Based on the rheotaxis behavior of sperm, a gradient flow field is created in the chip for progressive sperm sorting. Clinical experiment results for 10 volunteers showed that greater than 90% of isolated sperm exhibit high motility (> 25 μm/s), high linearity (0.8), and a very low DNA fragmentation rate (< 5%). In addition, the whole process is label and chemical free. These features aid in gentle sperm sorting to obtain healthy sperm. This device uniquely enables the selection of high-quality sperm with progressive motility and might be clinically applied for infertility treatment in the near future.

Keywords: Biodevices; Biotechnology; Interdisciplinary physics.

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Conflict of interest statement

The authors declare no competing interests.

Figures

None
Graphical abstract
Figure 1
Figure 1
Overview of the Progressive Sperm Sorting Chip (PSSC) (A) Illustration of the experimental setup. Both semen input and sperm are sorted by two syringe pumps, and the flow of mHTF buffer is driven by gravity with a Progressive Sperm Sorting Chip (PSSC). (B) Photo of the microfluidic chip with blue buffer in a microchannel. (C) Figure illustrating three different functional areas of PSSC. (D) COMSOL simulation results showing the velocity profile at the collecting zone of the microchannel. A low flow velocity occurs in the distal collector while loading the sample.
Figure 2
Figure 2
Model for the rheotaxis characteristic of human sperm (A) Images of cells moving in the retarding flow field. A Sperm velocity (V_s) over 25μm s-1 separated based on rheotaxis characteristic and a Sperm velocity (V_s) less than 25μm s-1. Cells unable to move can be washed to a backward outlet. (B–D) The relationship between linear velocity and linearity in sorted sperm.
Figure 3
Figure 3
Comparison of the sperm sorting efficiency methods for linear velocity and linearity (A) by liquefying the original semen sample, (B) using a PSSC device, (C) the swim-up method, and (D) gradient centrifugation.
Figure 4
Figure 4
Sperm morphology and DNA damage Comparison (A) of the figure shows the morphological characteristics of sperm, including (i) the original morphology and (ii) the morphology after PSSC sorting. It also displays (iii) the percentage of normal and abnormal sperm. Panel (B) of the figure focuses on sperm DNA fragmentation. (i) The original sample is shown, along with samples sorted using (ii) density gradient centrifugation, (iii) the swim-up method, and (iv) PSSC chip separation. (v) The figure also illustrates the percentage of sperm with DNA damage, represented by blue circles for healthy sperm and red circles for unhealthy sperm.
Figure 5
Figure 5
Clinical Patient Comparison (A) Top view of sperm. The rotation caused by shear leads to an upstream orientation. Fs, sperm propulsive force. The angular velocity of this rotation (ω) can be described by ω = dθ/dt. (B) straight-line velocity (VSL), (C) linearity (LIN), and (D) Progressive sperm ratio.
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