1. 从江香猪睾丸支持细胞分离培养与鉴定.
- Author
-
高艺, 黄泳, 冯贤辀, 王维勇, 徐永健, and 龚婷
- Subjects
- *
SERTOLI cells , *CELL suspensions , *CELL proliferation , *EPITHELIAL cells , *COLLAGENASES , *CYTOPLASM - Abstract
【Objective】 This study aimed to establish an effective method for isolation and culture of testicular Sertoli cells (SCs) from the Congjiang Xiang pig in vitro, provide cell material for the following reproduction of male Xiang pig, and provide reference for the isolation and cultivation of SCs from same type of Xiang pig. 【Method】 Using the combined enzyme digestion method of 0.1% collagenase Ⅳ and 0.25% trypsin-EDTA, testicular tissues were digested to isolate SCs. SCs were purified by differential adhesion method and cultured in the complete medium containing 10% fetal bovine serum. The morphological characteristics and growth curve of SCs from the Congjiang Xiang pig were observed, and SCs was identified by hematoxylin-eosin staining, RT-PCR and immunofluorescence staining. Identifying SCs by hematoxylin-eosin staining, RT-PCR and immunofluorescence staining.【Result】 A large number of spermatogenic epithelial cell suspensions could be obtained after combined enzyme digestion. The isolated SCs were round or oval, and the adherent cells after culturing 5-6 hours were triangular or irregular. SCs showed tight junction after 3-4 days, and subsequently, the cell proliferation rate decreased after 7-8 days and the growth curve of primary Sertoli cells was S-shaped. HE staining showed that the nucleus of SCs was dark purple and the cytoplasm was light red; RT-PCR showed that Sertoli cell specific genes WT1 and SOX9 were expressed, and the positive rate of GATA-4(a SC-specific maker) was more than 90% in immunofluorescence staining.【Conclusion】 Aculture method of testicular SCs of the Congjiang Xiang pig in vitro is successfully established. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF