新生大鼠性别鉴定及细胞培养液激素测定

Identification gender of neonatal rats and determination of sex hormone concentration in cultured astrocytes

目的研究新生雌雄大鼠的性别鉴定及培养细胞激素测定。方法采用大体形态学和HE染色直接观察性别,SRY基因PCR扩增技术及化学发光标记免疫分析法(CLIA方法)测定细胞培养液中雌二醇(E2)和睾酮(T)的浓度。结果 18只新生大鼠中直接用大体形态学方法进行性别鉴定,雌性排泄器与生殖器的距离为0.1~0.2 cm,雄性0.5 cm,HE染色中无明显性器官组织形态,PCR扩增SRY基因检测中8只雌性,10只雄性;CLIA方法检测星形胶质细胞培养液中雌二醇浓度含量:雄性(235.85±8.38)pg/ml,雌性(224.12±7.86)pg/ml,睾酮浓度含量:雄性(0.95±0.104)ng/ml,雌性(0.95±0.045)ng/ml。结论大体形态学测定及SRY基因PCR方法均可用于新生大鼠性别鉴定,培养不同性别细胞培养液中测定雌二醇浓度不同。提示在实验动物性别对实验结果可能有影响。在临床中应考虑性别差异对疾病,尤其是肿瘤的发生、发展及愈后的影响。

OBJECTIVE To study the sexing newborn male and female rats and hormone concentration in supernatant of the cultured cells.METHODS Gross morphology,HE staining and PCR amplification of SRY gene technology was used in this study.The estradiol(E2)and testosterone(T) concentrations in supernatant of the cultured cells were detected with CLIA method.RESULTS Eighteen newborn rats were sexed by gross morphology.The distance between the excretory organ and genital organ was0.1-0.2cm in female newborn rats and 0.5cm in male newborn rats.HE staining was failed to distinguish the gender.Eight female and 10 male newborn rats were detected by PCR methods.The result of gross morphology was consistent with the PCR method.Estradiol and testosterone concentrations in supernatant of cultured astrocytes were(235.85±8.38) pg/ml and(0.95±0.104) pg/ml in male newborn rats,(224.12±7.86)ng/ml and(0.95±0.045) ng/ml in female newborn rats respectively.CONCLUSION Gross morphology and SRY gene PCR method can be used for sexing newborn rats,culture of different sex determination of cell culture medium in different concentrations of estradiol.The Sex identification and hormone determination can attract the experimenter to the sex identification.The occurrence,development and prognosis of head and neck tumor may refer to the gender and hormone levels clinically.