title: Live staining and isolation of specific hormone-producing cells from rat anterior pituitary by cytochemistry with lectins and cholera toxin B subunit.
abstract: Anterior pituitary glands contain five types of hormone-producing cells. Distinguishing and isolating specific types of living cells are essential for studying their function. Although many such attempts have been made, the results have been disappointing. In the present study, we labeled specific types of living hormone-producing cells by using potential differences in sugar chains on the cell surfaces. Cytochemical analysis with lectins and cholera toxin B subunit revealed that PNA, S-WGA, and cholera toxin B subunit recognized sugar chains specific to prolactin cells, ACTH cells, and GH cells, respectively, and that UEA-I recognized most of prolactin cells and GH cells. Next, fluorescence-activated cell sorting was used to isolate GH cells labeled by fluoresceinated cholera toxin B. The purity of the GH cell fraction estimated by immunocytochemistry and quantitative real-time PCR for cell type-specific genes was more than 98%, which was higher than that reported in earlier studies, including those using transgenic animals. We conclude that cytochemistry with lectins and cholera toxin B subunit is a straightforward, acceptable method of isolating specific types of anterior pituitary cells and that the cells isolated by this method can serve as useful materials in the study of anterior pituitary cells.
authors.affiliation: Laboratory of Education, Jichi Medical University.
authors.forename: Motoshi
authors.initials: M
authors.lastname: Kikuchi
authors.forename: Kenji
authors.initials: K
authors.lastname: Kusumoto
authors.forename: Ken
authors.initials: K
authors.lastname: Fujiwara
authors.forename: Kozue
authors.initials: K
authors.lastname: Takahashi
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Tando
authors.forename: Takashi
authors.initials: T
authors.lastname: Yashiro