title: Establishment and culture optimization of a new type of pituitary immortalized cell line.
abstract: The pituitary gland is a center of the endocrine system that controls homeostasis in an organism by secreting various hormones. The glandular anterior pituitary consists of five different cell types, each expressing specific hormones. However, their regulation and the appropriate conditions for their in vitro culture are not well defined. Here, we report the immortalization of mouse pituitary cells by introducing TERT, E6, and E7 transgenes. The immortalized cell lines mainly expressed a thyrotroph-specific thyroid stimulating hormone beta (Tshb). After optimization of the culture conditions, these immortalized cells proliferated and maintained morphological characteristics similar to those of primary pituitary cells under sphere culture conditions in DMEM/F12 medium supplemented with N2, B27, basic FGF, and EGF. These cell lines responded to PKA or PKC pathway activators and induced the expression of Tshb mRNA. Moreover, transplantation of the immortalized cell line into subcutaneous regions and kidney capsules of mice further increased Tshb expression. These results suggest that immortalization of pituitary cells with TERT, E6, and E7 transgenes is a useful method for generating proliferating cells for the in vitro analysis of pituitary regulatory mechanisms.
authors.affiliation: Graduate School of Life and Environmental Sciences, The University of Tsukuba, Tsukuba, Ibaraki 305-8562, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Kokubu
authors.affiliation: Graduate School of Life and Environmental Sciences, The University of Tsukuba, Tsukuba, Ibaraki 305-8562, Japan; Life Science Center of TARA, The University of Tsukuba, Ibaraki-ken 305-8577, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Asashima
authors.affiliation: Graduate School of Life and Environmental Sciences, The University of Tsukuba, Tsukuba, Ibaraki 305-8562, Japan; Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8562, Japan. Electronic address: akikuri@hotmail.com.
authors.forename: Akira
authors.initials: A
authors.lastname: Kurisaki
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed; D017690:Cell Transplantation; D051379:Mice; D008813:Mice, Inbred ICR; D010902:Pituitary Gland / Q000166:cytology*
keywords: E6; E7; Immortalization; Pituitary; TERT