Isolation of PRRX1-positive adult pituitary stem/progenitor cells from the marginal cell layer of the mouse anterior lobe
Isolation of PRRX1-positive adult pituitary stem/progenitor cells from the marginal cell layer of the mouse anterior lobe
复制标题
DOI：
10.1016/j.scr.2021.102223
复制
发表时间：
2021-02-06
影响因子：
1.2
通讯作者：
Higuchi，Masashi
中科院分区：
文献类型：
--
作者：
Shintani，Aran;Higuchi，Masashi
我们最近提出，配对相关同源异型盒1（PRRX 1）和性别决定区Y盒2（SOX 2）双阳性（PRRX 1/SOX 2阳性）细胞是垂体干/祖细胞。本研究旨在分离位于垂体干/祖细胞龛中的PRRX 1阳性细胞。免疫组织化学分析显示，PRRX 1/SOX 2阳性细胞主要位于边缘细胞层（MCL）的成年小鼠垂体的生态位，但很少出现在实质生态位。二维培养的原代细胞从前叶（AL），包括MCL-龛，实现了有效的增殖PRRX 1/SOX 2阳性细胞与垂体干/祖细胞和龛标记物的高表达水平。相比之下，来自AL的原代细胞（不包括MCL-小生境）显示出有限的生长。当分离的PRRX 1/SOX 2阳性细胞和集群的实质生态位进行了比较免疫细胞化学，水通道蛋白5，标记的MCL生态位，仅在分离的细胞中检测到。三维培养分离的PRRX 1/SOX 2阳性细胞促进囊样球体的形成和向内分泌细胞的分化。因此，PRRX 1阳性的成体垂体干/祖细胞成功地从MCL-龛分离。本研究为分析脑垂体的细胞供应系统提供了有力的工具。
We recently suggested that paired-related homeobox 1 (PRRX1) and sex-determining region Y-box 2 (SOX2) double-positive (PRRX1/SOX2-positive) cells are pituitary stem/progenitor cells. The present study aimed to isolate PRRX1-positive cells located in the pituitary stem/progenitor cell niche. Immunohistochemical analysis revealed that PRRX1/SOX2-positive cells were located mainly in the marginal cell layer (MCL)-niche of the adult mouse pituitary gland, but were rarely present in the parenchymal-niche. Two-dimensional cultivation of primary cells from the anterior lobe (AL), including the MCL-niche, achieved efficient proliferation of PRRX1/SOX2-positive cells with high expression levels of pituitary stem/progenitor cell and niche markers. In contrast, primary cells from the AL, excluding the MCL-niche, showed limited growth. When isolated PRRX1/SOX2-positive cells and clusters of the parenchymal-niche were compared immunocytochemically, aquaporin 5, a marker of the MCL-niche, was detected only in isolated cells. Three-dimensional cultivation of isolated PRRX1/SOX2-positive cells promoted the formation of cystic-like spheroids and differentiation into endocrine cells. Thus, PRRX1-positive adult pituitary stem/progenitor cells were successfully isolated from the MCL-niche. The present study provides a powerful tool to analyze the cell supply system in the pituitary gland.