| 摘要: |
| 【目的】 探讨山羊腺垂体细胞的分离和培养方法，为其体外分泌特性研究奠定基础。【方法】 采集山羊腺垂体，分别用胰蛋白酶、胶原酶、胰蛋白酶和胶原酶混合、胰蛋白酶和透明质酸酶混合对其进行消化，所获细胞培养后采用免疫细胞化学法进行鉴定，评价细胞消化结果和腺垂体细胞的生长特性。【结果】 山羊腺垂体组织在37 ℃条件下，以1 g/L Ⅳ型胶原酶和1 g/L透明质酸酶混合消化2.5~3 h，消化效果较好；所得细胞在培养24 h开始贴壁，72 h后细胞主要呈多角形或梭形，连接成片，培养6 d左右细胞即可铺满培养器皿的细胞生长面。免疫细胞化学鉴定结果显示，FSH阳性细胞胞质呈蓝色，PRL阳性细胞胞质呈蓝褐色；放射免疫分析法测定表明，分离、培养的腺垂体细胞具有激素分泌活性。【结论】 采用筛选的消化和分离方法可获得山羊腺垂体细胞，所获腺垂体细胞体外培养体系,可用于体外研究腺垂体细胞的功能和内分泌调控作用。 |
| 关键词: 腺垂体细胞；免疫细胞化学 放射免疫分析；山羊 |
| DOI： |
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| 基金项目:陕西省科学技术研究发展计划（农业攻关）项目（2005K02-G4-1）；农业部农业结构调整重大技术研究专项（05-07-03B） |
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| Isolation,culture and identification of goat anterior pituitary cells |
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YIN Hui-yong,ZHAO Xiao-e,MA Bao-hua
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| Abstract: |
| 【Objective】 This paper focuses on the establishment of a method of the goat anterior pituitary cells in vitro .It can set a basis for the study of secretion characteristics in vitro.【Method】 The effects of different enzymes,including trypsin,collagenase Ⅳ,trypsin plus hyaluronidase,collagenase Ⅳ plus hyaluronidase on anterior pituitary in goat were discussed.Immunocytochemical staining identification was conducted and growing characters evaluated.【Result】 The results showed that the tissues presented best digestion by 1 g/L collagenase Ⅳ plus 1 g/L hyaluronidase,at 37 ℃ 2.5－3 h;anterior pituitary cells became adherent after 24 h of culture;the form of spindle or polygonal cells were found after 72 h of culture;they covered petri dish after 6 d of culture.Immunocytochemical staining showed that the cytoplasm of positive FSH cells showed blue color,and the cytoplasm of PRL positive cells showed blue brown color,radioimmunoassay showed secrete activity.【Conclusion】 The conclusion can be drawn that anterior pituitary cells can be obtained by the digestion and separation method suggested in this article.The system is a useful model for the study of function and endocrine regulation of goat anterior pituitary cells in vitro. |
| Key words: anterior pituitary cell immunocytochemical radioimmunoassay goat |
