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Rapid microfluidic sperm isolation from microtese samples in men with non-obstructive azoospermia

T. Jenkins, R. Samuel, A. Jafek, H. Feng, B. Gale, D.T. Carrell, J.M. Hotaling
P-360 Tuesday, October 31, 2017
DOI: http://dx.doi.org/10.1016/j.fertnstert.2017.07.733


Objective
To significantly improve efficiency of sperm recovery and decrease processing time of MicroTESE samples from men with non-obstructive azoospermia (NOA).

Design
We have developed and tested a microfluidic system consisting of two modules. The first module isolates sperm from other larger cells from the biopsied tissue. The second module enriches and concentrates sperm collected from the first module. We assessed the ability to identify sperm in samples processed through our device versus samples processed in the typical fashion following the microTESE procedure. We compared each process by the metric of sperm identified/period of time.

Materials and Methods
A total of 5 Testicular biopsy samples have been assessed to date. These samples were manually disrupted and exposed to collagenase for approximately 1-2 hours. The resulting single cell suspension was mixed thoroughly in Quinns media and a small portion was observed under oil on an inverted scope (this represents the currently performed manual procedure in many infertility clinics). Following this, 400 μl of the same sample was diluted to 800 μl and run through the isolation and enrichment modules sequentially and the output was placed under oil for observation. The number of cells observed and the time required to find them are both recorded.

Results
When using the microfluidic system we were able to identify more sperm in less time than in the traditionally used manual method. Specifically, we were able to identify 6.825 times more sperm in each ½ hour increment in our device than by the manual method.

Conclusions
We present a prototype microfluidic system capable of performing label-free separation and enrichment of sperm from digested testicular biopsies from NOA patients. In less than 20 minutes our system takes in an input of ∼500-1000 μl of digested biopsy tissue and utilizes inertial microfluidic physics and filtration to output ∼50-100 μl of enriched volume of sperm. Often, clinicians and labs spend tremendous time with each patient sample to isolate a few sperm from such biopsies. The presented system can significantly aid this tedious process, while ensuring efficient recovery of sperm.

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