Method for isolating pure bovine gonadotrophs from anterior pituitary usingmagnetic nanoparticles and anti-gonadotropin-releasing hormone receptorantibody

The Journal of Veterinary Medical Science

Hiroya KADOKAWA¹  Ashrafun NAHAR¹  Kiran PANDEY¹ 

[1] Joint Faculty of Veterinary Medicine, YamaguchiUniversity, Yoshida 1677–1, Yamaguchi-shi, Yamaguchi 753–8515,Japan

关键词: Easysep;    gonadotroph;    luteinizing hormone;    ruminant;   

DOI  :  10.1292/jvms.16-0157

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

No methods are currently available for rapidly isolating gonadotrophs from the anteriorpituitary (AP) in any species. We developed a method for preparing pure bovinegonadotrophs from a heterogeneous AP cell mixture by magnetic separation and our originalantibody against the N terminus of bovine gonadotropin-releasing hormone receptor (GnRHR).A bovine AP cell mixture was incubated with the anti-GnRHR antibody, anti-dextranantibody-conjugated secondary antibody and dextran-coated magnetic nanoparticles formagnetic isolation. Approximately 5.2 × 106 cells were isolated per AP ofJapanese Black heifers (26 months of age) and cultured, and confocal microscopy confirmedto be GnRHR- and luteinizing hormone-positive, corresponding to a purity of 100%.Approximately 44.5 µg of total protein was extracted from the puregonadotrophs per AP.

【 授权许可】

Unknown