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Abstract
BackgroundSome children who have survived cancer will be azoospermic in the future. Performing isolation and purification procedures for spermatogonial stem cells (SSC) is very critical. In this regard, performing the process of decontamination of cancerous cells is the initial step. The major objective of the present study is to separate the malignant EL4 cell line in mice and spermatogonial stem cells in vitro.MethodsThe spermatogonial stem cells of sixty neonatal mice were isolated, and the procedure of co-culturing was carried out by EL4 which were classified into 2 major groups: (1) the control group (co-culture in a growth medium) and (2) the group of co-cultured cells which were separated using the microfluidic device. The percentage of cells was assessed using flow cytometry technique and common laboratory technique of immunocytochemistry and finally was confirmed through the laboratory technique of reverse transcription-polymerase chain reaction (RT-PCR).ResultsThe actual percentage of EL4 and SSC after isolation was collected at two outlets: the outputs for the smaller outlet were 0.12% for SSC and 42.14% for EL4, while in the larger outlet, the outputs were 80.38% for SSC and 0.32% for EL4; in the control group, the percentages of cells were 21.44% for SSC and 23.28% for EL4 (based on t test (p ≤ 0.05)).ConclusionsThe present study demonstrates that the use of the microfluidic device is effective in separating cancer cells from spermatogonial stem cells.
Highlights
Some children who have survived cancer will be azoospermic in the future
We presented a new technique of cell sorting of spermatogonial stem cells from cancerous cells
EL4 tumor cell culture was performed in Dulbecco’s modified Eagle’s medium (DMEM)/F12 medium with FBS 2%, and the percentage of viable cells was about 80 ± 2.4%
Summary
Some children who have survived cancer will be azoospermic in the future. Performing isolation and purification procedures for spermatogonial stem cells (SSC) is very critical. The spermatogonium that are present in the testicles from birth are the precursors to the production of male sex cells The presence of these kinds of cells is very essential for performing the process of spermatogenesis. These cells are not capable of making the mature sperm before the puberty period due to the fact that they are dependent on hormonal stimuli [1]. About one third of children during their puberty period would experience a considerable decrease in the number of sperms or may face with the medical condition of azoospermia [2, 3]. This can jeopardize their quality of life [4]
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