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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. - Highlights: • Mouse pituitary cell lines were immortalized by introducing TERT, E6, and E7. • The immortalized cell linesmore » mainly expressed thyroid stimulating hormone beta. • The cell lines responded to PKA or PKC pathway activators, and induced Tshb.« less

Authors:
 [1];  [1];  [2];  [1];  [3]
  1. Graduate School of Life and Environmental Sciences, The University of Tsukuba, Tsukuba, Ibaraki 305-8562 (Japan)
  2. (Japan)
  3. (AIST), Tsukuba, Ibaraki 305-8562 (Japan)
Publication Date:
OSTI Identifier:
22462168
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 463; Journal Issue: 4; Other Information: Copyright (c) 2015 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CAPSULES; HOMEOSTASIS; IN VITRO; KIDNEYS; MESSENGER-RNA; MICE; OPTIMIZATION; PITUITARY GLAND; TSH

Citation Formats

Kokubu, Yuko, Asashima, Makoto, Life Science Center of TARA, The University of Tsukuba, Ibaraki-ken 305-8577, Kurisaki, Akira, E-mail: akikuri@hotmail.com, and Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology. Establishment and culture optimization of a new type of pituitary immortalized cell line. United States: N. p., 2015. Web. doi:10.1016/J.BBRC.2015.06.088.
Kokubu, Yuko, Asashima, Makoto, Life Science Center of TARA, The University of Tsukuba, Ibaraki-ken 305-8577, Kurisaki, Akira, E-mail: akikuri@hotmail.com, & Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology. Establishment and culture optimization of a new type of pituitary immortalized cell line. United States. doi:10.1016/J.BBRC.2015.06.088.
Kokubu, Yuko, Asashima, Makoto, Life Science Center of TARA, The University of Tsukuba, Ibaraki-ken 305-8577, Kurisaki, Akira, E-mail: akikuri@hotmail.com, and Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology. Fri . "Establishment and culture optimization of a new type of pituitary immortalized cell line". United States. doi:10.1016/J.BBRC.2015.06.088.
@article{osti_22462168,
title = {Establishment and culture optimization of a new type of pituitary immortalized cell line},
author = {Kokubu, Yuko and Asashima, Makoto and Life Science Center of TARA, The University of Tsukuba, Ibaraki-ken 305-8577 and Kurisaki, Akira, E-mail: akikuri@hotmail.com and Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology},
abstractNote = {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. - Highlights: • Mouse pituitary cell lines were immortalized by introducing TERT, E6, and E7. • The immortalized cell lines mainly expressed thyroid stimulating hormone beta. • The cell lines responded to PKA or PKC pathway activators, and induced Tshb.},
doi = {10.1016/J.BBRC.2015.06.088},
journal = {Biochemical and Biophysical Research Communications},
number = 4,
volume = 463,
place = {United States},
year = {Fri Aug 07 00:00:00 EDT 2015},
month = {Fri Aug 07 00:00:00 EDT 2015}
}