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[目的]通过对大鼠前体精原干细胞-支持细胞共培养系进行镉染毒,探讨镉对前体精原干细胞和支持细胞共培养系早期效应的形态学变化。[方法]从3天龄的SD大鼠睾丸中分离前体精原干细胞和支持细胞并进行24h共培养后,再分别给予0、2.5、5.0、10.0、20.0μmol/L的氯化镉(CdCl2)染毒12h,然后用碱性磷酸酶和油红O染色分别鉴定前体精原干细胞和支持细胞,观察不同细胞的形态,并用原位末端标识法(TUNEL方法)进行细胞凋亡的原位检测。[结果]2.5μmol/LCdCl2染毒未显著改变共培养中精原干细胞和支持细胞的形态。5.0μmol/L以上浓度CdCl2染毒可致支持细胞(由油红O染色确认)的结构呈现明显变化,具有浓度依赖趋势,用TUNEL法检测发现前体精原干细胞和支持细胞均发生凋亡,和对照组相比差别有统计学意义。[结论]CdCl2染毒12h可诱导支持细胞的形态改变,以及前体精原干细胞和支持细胞的凋亡。
[Objective] The purpose of this study was to investigate the morphological changes of early-phase effects of cadmium on the co-culture of progenitor spermatogonial stem cells and supporting cells by cadmium exposure of rat precursor spermatogonial stem cells-supporting cells co-culture system. [Method] Pre-cultured spermatogonial stem cells and supporting cells were isolated from 3-day-old SD rat testis and co-cultured for 24 hours. Then they were treated with 0,2.5,5.0,10.0 and 20.0μmol / L cadmium chloride ) For 12 hours. Then the spermatogonial stem cells and supporting cells were identified by alkaline phosphatase and oil red O staining. The morphological changes of different cells were observed, and the cell apoptosis was detected by TUNEL method Detection. [Result] The morphological changes of spermatogonial stem cells and supporting cells in co-culture were not significantly affected by 2.5 μmol / L CdCl2. The concentration of 5.0μmol / L CdCl2 exposed cells (identified by oil red O staining) showed obvious changes in the structure, with a concentration-dependent trend, detected by TUNEL method found that the precursor spermatogonial stem cells and supporting cells were apoptosis, Compared with the control group, the difference was statistically significant. [Conclusion] The morphological changes of supporting cells induced by CdCl2 for 12 hours and the apoptosis of precursor spermatogonial stem cells and supporting cells were observed.