镉致大鼠前体精原干细胞-支持细胞共培养系损伤的形态变化

Morphological Change Induced by Cadmium in Primary Rat Gonocyte Cell-Sertoli Co-cultures in vitro

  • 摘要:
    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24 h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12 h,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/L CdCl2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12 h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。

     

    Abstract:
    Objective To investigate the acute effects of cadmium on primary rat gonocyte cell/sertoli co-cultures.

    Methods Gonocytes and sertoli cells were separated from 3-day SD rat. Cadmium (0, 2.5, 5, 10, 20 μmol/L)were added to coculture system 12 h after gonocyte and sertoli cell had co-cultured for 24 h. Alkaline phosphatase and oilred O were used to identify the gonocyte and sertoli cells. TUNEL in situ detection was used for detecting apoptosis.

    Results Exposure of 2.5 μmol/L cadmium did not induce morphologic change of gonocytes and sertoli cells. When exposure dose greater than 5 μmol/L, sertoli cells showed morphological change identified by oilred O in a dose-dependent manner. TUNEL in situ detection suggested that cadmium in creased apoptosis in both gonocytes and sertoli cells.

    Conclusion Exposure of cadmium for 12 h induced morphological change of sertoli cells and apoptosis of both gonocyte and sertoli cell.

     

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