The invention relates to the field of
molecular biology and specifically discloses an immortalized rabbit
small intestine epithelium cell line and a construction method thereof. The small intestines of a newly born rabbit are collected, the
enzymatic digestion juice containing
collagenase and
neutral protease is utilized to digest, and after the
digestion, a method for re-suspending centrifuging and differential adherence is adopted for purifying the pit
cell cluster, so that high-purity primary pit
epithelium cells can be obtained. The pit
epithelium cells can participate in culture and passage. The obtained pit epithelium cells are used for performing
liposome transfection expression on the SV40 big T antigenic
plasmid, the passage is continued, and the immortalized rabbit
small intestine epithelium
cell line is finally screened. The immortalized rabbit
small intestine epithelium cell line can simultaneously express intestine epithelium
surface marker molecules and pit basic
stem cell surface molecules. The intestine epithelium
surface marker molecules include but not limited to
keratin 8, E-
calcitonin and ki67; and the pit basic
stem cell surface molecules include but not limited to Lgr5 and sox9. The immortalized rabbit small intestine epithelium cell line can be used for establishing a rabbit small intestine epithelium
in vitro model and can be used for researching a
nutrition absorbing mechanism, cellular pathway excavation and
pathogen in vitro culture.