Shahdad Ronak Commercialization Company, Pasdaran Street, Tehran, Iran.
Radiation Biology Research Center, Iran University of Medical Sciences, Tehran, Iran.
Stem Cell Res Ther. 2020 May 24;11(1):191. doi: 10.1186/s13287-020-01671-1.
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. 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.
The 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).
The 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)).
The present study demonstrates that the use of the microfluidic device is effective in separating cancer cells from spermatogonial stem cells.
一些患有癌症的儿童将来会不育。分离和纯化精原干细胞（SSC）的程序非常关键。在这方面，进行癌细胞的净化过程是初始步骤。本研究的主要目的是在体外分离小鼠中的恶性 EL4 细胞系和精原干细胞。
从 60 只新生小鼠中分离出精原干细胞，并通过 EL4 进行共培养，分为 2 个主要组：（1）对照组（在生长培养基中进行共培养）和（2）使用微流控设备分离的共培养细胞组。使用流式细胞术技术评估细胞百分比，并通过免疫细胞化学的常见实验室技术进行最后确认，最终通过逆转录-聚合酶链反应（RT-PCR）的实验室技术进行确认。
在两个出口处收集分离后的 EL4 和 SSC 的实际百分比：较小出口的输出为 SSC 的 0.12%和 EL4 的 42.14%，而较大出口的输出为 SSC 的 80.38%和 EL4 的 0.32%；在对照组中，SSC 的细胞百分比为 21.44%，EL4 的细胞百分比为 23.28%（基于 t 检验（p≤0.05））。
本研究表明，使用微流控设备可有效分离癌细胞和精原干细胞。
