The invention discloses a method for constructing a machin
progenitor leydig cell immortal line. The method comprises the following steps: by taking machin testicular tissues, obtaining leydig cells of a
high ratio, culturing in a complete medium for 20-28 hours, replacing fresh culture solution, continuously culturing, and performing
cell passage to the leydig cells presenting
fibroblast growth in uniform form; infecting the leydig cells by utilizing
slow virus solution carrying Large T and Egfp, performing
fluorescence labeling
cell screening, and performing
cell passage to a
cell strain for stably expressing Egfp; and screening the
cell strain for stably expressing Egfp, identifying the
progenitor leydig cells, continuously performing cell passage for over 50 generations, and finally constructing the machin
progenitor leydig cell immortal line. The in-vitro cell passage of the progenitor
leydig cell immortal line with the transferred Large T exceeds 62 generations, the leydig cell labeled proteins such as CYP11A1, CYP17A1 and STAR are expressed, marker genes of mesenchymal stem cells such as Pdgfral and Lifr are expressed, the
cell shape is maintained into the
fiber shape, and a characteristic progenitor leydig cell immortal line is obtained.